Abstract:
The invention relates to a device for obtaining biological material and/or biological information from a sample with a heterogeneous matrix, to the uses of said device, to methods implementing said device, and to kits for obtaining biological material and/or biological information from a sample with a heterogeneous matrix, comprising the different constituents of said device.

Description:
TECHNICAL FIELD 
       [0001]    The present invention relates to the field of obtaining biological material and/or biological information from a sample with a heterogeneous matrix for the purposes of subsequent analyses, in particular for diagnostic purposes. 
       PRIOR ART 
       [0002]    The testing of biological material and/or biological information from samples of various origins (for example of industrial, food-processing or clinical origin) requires the implementation of techniques which allow detection—for example for the purposes of identifying and/or counting microorganisms—and the yield of which in terms of results must be as efficient as possible. Generally, if said biological material is considered, the latter may be of non-pathogenic nature. However, in the diagnostic field, the biological material which is the subject of testing is usually pathogenic, consequently requiring rapid and precise detection thereof in order to initiate corrective actions ad hoc as soon as possible. Quite obviously, the toxins and other metabolites produced by this pathogenic biological material can also be investigated. 
         [0003]    In standard fashion, the detection of biological material and/or biological information can be carried out by means of immunological assays, enzymatic assays, molecular biology techniques, or else, as regards said biological material, by counting and/or identification on a culture dish. In any event, whether it is a question of biological material and/or of biological information, the detection sensitivity and the detection specificity are two important factors for evaluating the efficiency of the test performed. 
         [0004]    By way of example, the detection of microorganisms is conventionally carried out on selective culture media plated out on a Petri dish, as recommended by the standards of ISO (International Organization for Standardization) type or the BAM-FDA (Bacteriological Analytical Manual of the Food and Drug Administration) methods. These standards recommend in particular the use of media such as polymyxin egg yolk mannitol bromothymol blue Agar (the acronym of which is PEMBA) or mannitol-egg yolk-polymyxin Agar (the acronym of which is MYP). The identification of microorganisms is conventionally carried out according to morphological, culture and/or metabolic criteria. Nevertheless, these methods of detection on a Petri dish can be negatively affected by the presence of non-specific elements, derived from the sample to be analyzed. This is particularly true with regard to samples with a heterogeneous matrix, namely samples which have differences in terms of consistency (solid particles, parts that are more or less liquid, semi-solid parts, etc.), such as soil, human or animal stool, phlegm or sputum samples or else tissue samples (medico-legal samples), which are generally rich in polysaccharides, cell debris, and inhibitors of all types (bile salt, polyphenols, polysaccharide, fibers, hemoglobin, etc.), capable of disrupting the growth of the microorganisms sought and thus of giving rise to false-negative results (detection sensitivity problem). 
         [0005]    Alternatively, the detection of biological information derived from the biological material can be carried out, for example, using conventional methods for amplifying nucleic acids, specific for one or more types of microorganisms and viruses that are sought. However, these techniques are sensitive and may be significantly inhibited by compounds such as polysaccharides, cell debris and inhibitors of all types. This can result in the obtaining of false-negative results (detection sensitivity problems) or in an incorrect diagnosis and can have serious consequences for the human or animal patient. Consequently, it is essential to isolate the microorganisms and/or the viruses such that a minimum number of inhibitors are present in the isolate. This is particularly important for samples with a heterogeneous matrix, as previously mentioned. This is because said samples comprise a large number of non-specific constituents (polysaccharides, cell debris, inhibitors of all types), capable of generating false negatives during the implementation of the abovementioned nucleic acid amplification methods. 
         [0006]    In addition, and regardless of the type of detection method used, these samples with a heterogeneous matrix have, in addition to the conventional problems of contamination of the sample and/or of the environment, more specific problems with respect to metering out/calibration (correlation difficult between amount of the sample taken and mass thereof, means of calibration constant from one sample to the other) and with respect to suspending in a liquid medium, that are inherent in the nature of the sample. 
         [0007]    Currently, various methods for analyzing the genetic material contained in a sample with a heterogeneous matrix exist. By way of example, the “QIAamp® DNA” kit from the company QiaGen makes it possible to isolate and purify the total genomic DNA of a solid or liquid sample (such as blood, serum, plasma, etc.). This kit uses a device consisting of filtration columns and tubes suitable for use with standard microcentrifuges. According to the supplier&#39;s recommendations, the method for obtaining the total genomic DNA comprises the following steps: taking a given amount of the sample of interest (by weighing) depending on its nature, introducing said sample into a tube containing a solution for suspending it, placing the sample in suspension by means of vigorous stirring of vortex type. Once the sample has been vigorously mixed, a lysis solution is added and an incubation step is carried out for several minutes, in order to break the cell membranes and to release the genetic information into the solution. Next, in order to remove the debris, the solution is centrifuged and the supernatant is removed. There are subsequently laborious steps of adding solutions and performing centrifugations which make it possible to purify the DNA sample, before eluting it on a column by centrifugation. 
         [0008]    However, this method, using the abovementioned “QIAamp® DNA” kit, has several drawbacks, in particular when the sample tested is a sample with a heterogeneous matrix. This is because the first steps of weighing and transferring the sample into the microcentrifuge tubes are critical owing to the narrowness of the opening of said tubes. This transfer can thus prove to be difficult and can require additional handling and equipment, and extended implementation times. In addition, during these weighing and/or transfer operations, there is a significant risk of:
       loss of the material,   contamination of the sample and/or of the environment and/or of the laboratory technician,   discomfort of the laboratory technician associated with the repeated handling of samples of stool, sputum or phlegm type,   difficulty in calibrating the sample by weighing.       
 
         [0013]    Moreover, the numerous steps constituting the protocol of the “QIAamp® DNA” kit, which involve the handling of a considerable amount of different tubes, are capable of causing handling errors. Furthermore, the handling of small tubes, such as tubes suitable for microcentrifuges, does not prove to be very practical for the laboratory technician, and is therefore also capable of giving rise to losses in terms of time and efficiency. In addition, the “QIAamp® DNA” kit enables only the isolation of nucleic acid from a sample, and does not make it possible to detect the presence of viable microorganisms, for instance detection on culture medium. Furthermore, the “QIAamp® DNA” kit does not enable the detection of yeast from a stool sample. 
         [0014]    In another example, patent U.S. Pat. No. 4,081,356 describes a device and a method for recovering small sediments containing parasite eggs and larvae from a sample of fecal material. The device disclosed consists of two compartments separated by a basic (namely not very selective) filtration element, such as a filter and a steel mesh; the upper compartment being obstructed at its external end by a spatula enabling the taking of a sample. The sample of fecal material is taken by means of the spatula, which is suitable for being connected to the upper part of the compartment. Next, the sample is emulsified using a solution of formaldehyde, optionally supplemented with a surfactant, mechanically by means of a rod. The emulsification is finalized by a step of horizontal agitation in the presence of glass beads. A vertical agitation step then makes it possible to pass the entire emulsified sample into the lower compartment of the device, through the basic filtration element. Ether is then used to rinse this basic filtration element and to allow the formation of a mixture in the lower compartment. Finally, the lower compartment is agitated and centrifuged to separate the resulting mixture into various layers of materials of different densities, in order to isolate the layer of sediment containing the parasite eggs and the larvae. 
         [0015]    However, the device used in the abovementioned method has several drawbacks, in particular associated with the flagrant lack of selectivity of the basic filtration element, which allows the passage, in addition to the abovementioned eggs and larvae, of non-specific elements that will subsequently sediment in various layers. Moreover, the first steps of taking the sample, in particular the step of connecting the spatula to the upper compartment, do not allow clean and easy taking of a sample and, consequently, require additional handlings and extended implementation times. Furthermore, this inevitably affects the reproducibility of the method using this device, insofar as it proves to be extremely difficult—or even impossible—to take a precise and calibrated volume of said sample, in particular from hard stools of type 1 and 2 on the Bristol scale. Human stools are commonly characterized and classified according to the Bristol scale consisting of a visual classification which divides human stools up into seven types. Types 1-2 correspond to hard stools, types 3-4 are considered to be normal, and types 5-6-7 correspond to stools of liquid consistency. 
         [0016]    In view of the problems presented above, an objective of the present invention aims to develop a device and the process(es) associated therewith making it possible:
       to meter out/calibrate the amount of sample with a heterogeneous matrix that is taken without needing to weigh the latter,   to efficiently suspend said sample in a liquid medium,   to limit the problems associated with clogging (obstruction) of the filters in order to facilitate filtration and to improve recovery of the biological material and/or of the biological information,   to limit the obtaining of non-specific elements capable of distorting the result of the analysis in fine,   to prevent the loss of a part of the sample during the sample transfer into the container (for example into the tube),   to improve the efficiency and practicality,   to limit as much as possible the risks of contamination of the sample and/or of the environment and/or of the laboratory technician,   to simplify the protocol for preparing samples with a heterogeneous matrix in order to reduce the operating time, the number of materials and consumables used, and the risks of errors and to improve the reproducibility, to propose a device compatible with all of the known analytical methods (microbiology, culture, virology, bacteriology, immunoassay, metasequencing, PCR, etc.),   to isolate yeasts.       
 
         [0026]    Other objectives will emerge, as appropriate, on reading the present patent application. 
       SUMMARY OF THE INVENTION 
       [0027]    Consequently, the subject of the present invention relates to a device for obtaining biological material and/or biological information from a sample with a heterogeneous matrix, for example a human or animal stool (feces) sample, said device comprising:
       a container suitable for receiving a content comprising said sample and at least one suspending solution, intended to enable said sample to be placed in suspension,   a stopper which makes it possible to close said container, preferably hermetically, said stopper comprising:   at least one calibrated sampling means, which makes it possible to take a predetermined volume of said sample corresponding to a given mass, said sampling means comprising at least one calibrated hollow part connected to the internal part of the stopper and extending from said internal part of the stopper to the inside of the container when said stopper and said container are assembled,   at least one opening, which can preferably be closed by a removable closure means, allowing communication of fluid between the interior of said container and the exterior of said device when said stopper and said container are assembled,   at least one filtration means, positioned with respect to said opening in such a way as to filter the content while said content passes from the interior of said container to the exterior of said device via said opening, said filtration means being suitable for allowing the selective passage of said biological material and/or biological information to the exterior of said device.       
 
         [0033]    Thus, the device according to the invention makes it possible to simplify and facilitate the taking of a given mass of a sample with a heterogeneous matrix (in particular while dispensing with a difficult weighing step), the placing of said sample in suspension and the isolation of the biological material and/or of the biological information that it contains, with a view to the subsequent analysis thereof. In particular, the device according to the invention has the advantage of being suitable for treating samples with a heterogeneous matrix having a very varied consistency and/or texture. 
         [0034]    The simplification of the isolation of the biological material and/or of the biological information obtained by virtue of the device according to the invention leads in particular to a decrease in the “open tube” steps, which require consumables and technical manipulations (pipetting, removal of supernatant, centrifugation, etc.), capable of generating errors and contaminations of the sample and/or of the outside environment. The device according to the invention also makes it possible to dispense with the use of multiple pieces of equipment, such as balances, bead beaters or centrifuges. 
         [0035]    The term “sample with a heterogeneous matrix” is intended to mean a small part or a small amount isolated from a material with a heterogeneous matrix, for analysis. The material with a heterogeneous matrix from which the sample is taken is characterized by intrinsic differences in terms of consistency and/or of texture (solid parts, parts that are more or less liquid, semi-solid parts, viscous/viscoelastic parts). This material with a heterogeneous matrix is capable of containing the biological material and/or the biological information sought. By way of example, the sample with a heterogeneous matrix may be a soil, human or animal stool, phlegm or sputum sample, an industrial sample or else a tissue sample (medico-legal samples). Preferably, the sample with a heterogeneous matrix is a human or animal stool sample. According to one particularly preferred embodiment, this sample with a heterogeneous matrix is a human stool sample. The device according to the present invention makes it possible to obtain biological material and/or biological information from all of the types of stools defined by the Bristol scale, namely types 1-7. 
         [0036]    For the purposes of the present invention, the term “biological material” is intended to mean any material containing genetic information and which is self-reproducible or reproducible in a biological system. By way of example of the self-reproducible biological material, mention may be made of a microscopic self-reproducible “biological material”, such as one or more human, animal or plant cell(s) or one/several bacteria/yeasts. 
         [0037]    According to one preferred embodiment of the invention, said biological material is a self-reproducible microbiological material (such as one or more bacterium or bacteria, yeast or yeasts, fungus or fungi) or a biological material (such as one or more viruses) that is reproducible in a biological system. The device according to the present invention is particularly suitable for detecting and/or identifying and/or counting microorganisms that are pathogenic to humans and/or animals. 
         [0038]    For the purposes of the present invention, the term “biological information” is intended to mean any element constituting said biological material or produced by the latter, such as nucleic acids (DNA, RNA), proteins, peptides or metabolites. The biological information can in particular be contained within said biological material or excreted/secreted by the latter. 
         [0039]    Said “calibrated hollow part”, preferably of concave shape, makes it possible to delimit a predetermined volume, suitable for sampling a predefined mass of the material with a heterogeneous matrix. More specifically, the volume of this hollow part is determined so as to contain an amount of biological material and/or of biological information that is sufficient to allow the analysis thereof. 
         [0040]    According to one preferred embodiment, the calibrated sampling means comprises a rod and a calibrated hollow part. 
         [0041]    According to this preferred embodiment, said calibrated hollow part is connected to the internal part of the stopper by means of said rod. 
         [0042]    Advantageously, in order to avoid sampling an excess mass/amount of biological material and/or of biological information, said rod is adapted so that only said at least one calibrated hollow part is capable of containing sample with a heterogeneous matrix. To this effect, said rod is preferably devoid of any recess or cavity, obviously with the exception of said calibrated hollow part. According to one particular embodiment, said rod is made of solid material. 
         [0043]    According to one preferred embodiment, said calibrated hollow part is a hollowed-out part of said rod, advantageously located at the level of the end of the rod opposite the internal part of the stopper. 
         [0044]    According to one preferred embodiment, said hollow part has at least one wall suitable for removing the surplus sample with a heterogeneous matrix by “scraping” said calibrated hollow part against an appropriate surface, of sufficiently rigid constitution. Thus, by removing the surplus sample with a heterogeneous matrix, protruding outside the volume defined by the calibrated hollow part, the operator is sure that the mass of sample with a heterogeneous matrix removed does not significantly exceed the desired mass, corresponding to the volume of the calibrated hollow part. 
         [0045]    Advantageously, said hollow part is calibrated so as to contain a mass of between 10 mg and 2000 mg, preferably between 100 mg and 1000 mg, advantageously between 200 mg and 300 mg, of said material with a heterogeneous matrix. 
         [0046]    According to one particular embodiment of the invention, said rod comprises—or is connected to—at least one sliding part, making it possible to pass from a first length of rod to a second length of rod (and, preferably, conversely, namely to pass from said second length of rod to said first length of rod), said second length of rod being less than said first length of rod. 
         [0047]    According to a first aspect of this particular embodiment, said rod may be—at least partially—telescopic. 
         [0048]    According to a second aspect of this particular embodiment, that is particularly advantageous, the abovementioned sliding part is a sliding extension, suitable for being positioned on said rod and for sliding along the latter in order to pass from said first length of rod to said second length of rod (and, preferably, conversely), said sliding extension comprising at least one calibrated hollow part, preferably positioned at the end of the sliding extension that is furthest from the internal part of the stopper. 
         [0049]    According to one particular embodiment of the invention, the sampling means is suitable for containing several calibrated hollow parts, preferably from 2 to 10, preferentially from 2 to 6, advantageously 3 or 6, which can each contain between 10 mg and 2000 mg of said material with a heterogeneous matrix, preferably between 100 mg and 1000 mg, advantageously between 200 mg and 300 mg, of said material with a heterogeneous matrix. Consequently, according to this particular embodiment, the sampling means according to the invention can be suitable for taking an amount of sample with a heterogeneous matrix of between 20 mg and 20 000 mg, preferentially between 200 mg and 6000 mg. Obviously, those skilled in the art will know how to adapt the sampling means according to the invention, in particular with regard to the number and/or the volume of the calibrated hollow part(s), in particular in order to obtain, in fine, a concentration of biological material and/or biological information that is compatible with the minimum level of sensitivity of the analytical techniques. By way of example, it is well known to those skilled in the art that the microorganisms responsible for bovine tuberculosis (mycobacteria of the tuberculosis complex) are difficult to detect and consequently require a larger amount of sample in order to obtain an analyzable concentration of microorganisms. 
         [0050]    In one particular embodiment, said sampling means has, on each side of the rod, three calibrated hollow parts that can each contain a volume corresponding to 1000 mg, i.e. a total volume corresponding to 6000 mg, of sample with a heterogeneous matrix, such as bovine, ovine, caprine or porcine feces, advantageously bovine feces. 
         [0051]    Typically, the analysis of stools of human origin is carried out using from 200 to 300 mg of sample. However, in certain particular cases, this can range up to 1000 mg. Moreover, in the case of veterinary applications, the mass of sample required for the purposes of the analysis can represent several grams, or even several tens of grams. 
         [0052]    According to one preferred embodiment of the invention, the assembly of the stopper and container operates by tight cooperation of the two entities via a closure system, such as a screw-fastening or clip-fastening system, making it possible to close said container such that the content can only leave in the form of filtrate via said at least one opening of said stopper when said container is closed by said stopper. 
         [0053]    For the purposes of the present invention, the term “removable closure means” is intended to mean any means which makes it possible to prevent any uncontrolled exit of filtrate. By way of example, said closure means may be a cap or a protective breakable part. In the latter case, said protective breakable part is rigidly connected to said opening, preferably is sealed to the latter (for example by heat sealing) and proves to be breakable, for example by twisting or tensile force. Alternatively, this protective breakable part may be cut via the action of a slicing means, such as a knife or a pair of scissors. It is interesting to note that said breakable part also performs the function of impregnability control, since the fact that this breakable part is separated from the device means, for the operator, that the device in question has already been used. In other words, this breakable part makes it possible to ensure the integrity of the device according to the invention. 
         [0054]    Advantageously, when the removable closure means according to the invention is a protective breakable part, the latter has, on the end opposite said opening, a shape complementary to said opening. Thus, when said protective breakable part is separated from said opening, for example by applying a tensile or twisted force to said breakable part, the end of this breakable part having a shape complementary to said opening can be positioned on the latter in order to close it. In this preferred embodiment, said protective breakable part, once separated from the opening, acts as a cap; it thus being possible to describe said protective breakable part as a “protective breakable cap”. 
         [0055]    As previously indicated, the calibrated sampling means according to the invention is connected to the internal part of the stopper by any appropriate means. Thus, said calibrated sampling means can be screwed, adhesively bonded, assembled by elastic interlocking (for example by clip-fastening) or else forcibly plugged into an opening made in said stopper. According to one particular embodiment, this calibrated sampling means may be heat-sealed to the internal part of the stopper. 
         [0056]    According to one preferred embodiment of the invention, said container also comprises at least one mechanical suspending means, which is preferably spherical in shape, such as a bead, suitable for facilitating the suspension of said sample in said suspending solution. 
         [0057]    The term “mechanical suspending means” is intended to mean an element suitable for being placed within the container according to the invention and for being moved by one or more force system(s) external to the device according to the invention, for example by manual agitation or agitation induced by the mixing apparatus of the type such as a vortex, sonicator or magnetic stirrer, or bead beater in order to enable/facilitate the mechanical suspending of the sample within said suspending solution. 
         [0058]    Advantageously, the device comprises a number of mechanical suspending means, such as beads, selected between 1 and 200, preferably between 5 and 50, advantageously between 10 and 40, particularly preferably between 25 and 35; said mechanical suspending mean(s) having a size of between 1 mm and 10 mm, preferably between 2.5 mm and 3.5 mm, advantageously of approximately 3 mm; preferably, said mechanical suspending mean(s) being made of glass, iron, plastic(s), ceramic(s), particularly preferably of glass. 
         [0059]    The size of the mechanical suspending mean(s) is understood to be the largest dimension of the latter. For example, the size of a mechanical suspending means having the shape of a rectangular parallelepiped (straight block) is understood to mean the length (largest dimension) of this rectangular parallelepiped. As previously indicated, said at least one mechanical suspending means is spherical in shape and advantageously consists of a bead. In the case of a mechanical suspending means that is spherical in shape, the size is understood to be the diameter thereof. Thus, when use is made, as mechanical suspending means, of a plurality of beads having a size of between 2 mm and 10 mm, preferably between 2.5 mm and 3.5 mm, advantageously of approximately 3 mm, it should be understood that the diameter of the beads is between 2 mm and 10 mm, preferably between 2.5 mm and 3.5 mm, said beads advantageously having a diameter of approximately 3 mm. Preferentially, said size of said beads is in agreement with the shape and the dimensions of said calibrated hollow part. 
         [0060]    Preferably, the shape and/or the size of said at least one mechanical suspending means is/are suitable so as not to close said at least one opening allowing communication of fluid between the interior of said container and the exterior of said device when said stopper and said container are assembled. Conversely, when the size and/or the shape of said at least one mechanical suspending means is known in advance, the shape and/or the size of said opening can be adapted accordingly, in order to achieve the same objective. 
         [0061]    According to one particular embodiment, the mechanical suspending mean(s) has (have) a size (a diameter in the context of mechanical suspending mean(s) of spherical shape) suitable for cooperating with said at least one hollow part of the calibrated sampling means, in particular in order to facilitate the suspending of the sample with a heterogeneous matrix previously taken. 
         [0062]    Preferentially, the mechanical suspending mean(s) consist(s) of a material suitable for interacting with an external suspending system/device, said system/device making it possible to move or vibrate the mechanical suspending mean(s) (preferably the beads). Such a system/device may consist, for example, of a sonicator or a magnetic stirrer. Optimally, the material of which the mechanical suspending mean(s) consist(s) interacts little or not at all with said biological material and/or biological information, in particular so as not to modify/degrade said biological material and/or said biological information. 
         [0063]    Advantageously, in order to facilitate handling, said calibrated sampling means has sufficient rigidity to prevent said calibrated sampling means from curving while taking sample and to guarantee a low coefficient of variation with respect to the mass of biological matrix sampled. 
         [0064]    The term “sufficient rigidity” is intended to mean a rigidity suitable for taking all types of samples with a heterogeneous matrix, and in particular those which have a relatively solid consistency. The sufficient rigidity—and consequently the low elasticity—of the calibrated sampling means makes it possible to facilitate the sampling and to prevent projections of sample with a heterogeneous matrix during the sampling operation. 
         [0065]    Advantageously, the sufficient rigidity of the sampling means is obtained by using materials which make it possible to achieve the required degree of rigidity, such as polypropylene (PP), Delrin resin, polyamide (PA) thermoplastic materials, polycarbonate (PC), poly(methyl methacrylate) (PMMA), low-density polyethylene (LDPE), acrylonitrile-butadiene-styrene (ABS) and/or by means of a form which promotes the rigidity of the sampling means. In addition, in order to prevent the calibrated sampling means according to the invention from curving while taking sample, it proves to be important for the connection between said sampling means and the internal part of the stopper to be sufficiently rigid in order to prevent it from folding/curving during the sample-taking operation. Indeed, during this operation, the operator generally holds the external part of the stopper between at least two of his fingers and then takes the desired sample. It is thus important for there to be no weak point at the level of the connection between, on the one hand, the internal part of the stopper and, on the other hand, the calibrated sampling means. 
         [0066]    According to one particular embodiment, said calibrated hollow part comprises at least one opening which makes it possible to facilitate the suspending of said sample in said suspending solution. 
         [0067]    Preferably, said at least one opening has a circular, ovoid or elongated (preferably oblong) shape. Preferably, said opening is elongated, advantageously oblong, in shape, having for example a stick shape. This elongated, and preferably oblong, shape makes it possible to ensure calibrated and reproducible taking of the sample with a heterogeneous matrix while at the same time promoting the suspending thereof in the suspending solution. According to one particularly preferred embodiment, said at least one opening has a shape and/or a size suitable for cooperating with at least one mechanical suspending means as previously defined, in order to promote the suspending of the sample with a heterogeneous matrix taken by the operator. 
         [0068]    According to one particular embodiment, in order to further facilitate this suspending, said at least one hollow part is coated over all or part of its surface (preferably over all of its surface) with a non-stick coating, such as one or more layer(s) of water-soluble material(s) or else a coating of hydrophobic type. Preferably, and in order to facilitate the manufacture of the device, the material chosen for producing the calibrated hollow part has a natural tendency to be hydrophobic and/or exhibits low roughness. By way of example, this low roughness may in particular be obtained by polishing so as to give a polished mirror finish. 
         [0069]    According to one preferred embodiment, said container comprises said suspending solution, for example a buffer solution, in a volume sufficient to allow the suspending of the sample. 
         [0070]    This suspending can be carried out:
       directly, namely by partially or totally, preferably totally, immersing said at least one calibrated hollow part containing the sample in the suspending solution, or   indirectly, by inducing a variation in level of the suspending solution, such that the latter comes into intermittent contact (at regular or irregular intervals) with all or part of said at least one calibrated hollow part containing the sample.       
 
         [0073]    The “indirect” suspending of the sample with a heterogeneous matrix in the suspending solution can in particular occur during a mechanical suspending step, namely, for example, during a stirring/mixing step. The latter can be obtained, for example, by using a stirrer or a mixing apparatus of vortex type. 
         [0074]    By way of example of a “suspending solution” it is possible to use any solution suitable for:
       i) allowing the suspending of said “sample with a heterogeneous matrix”, and/or   ii) preserving the biological matrix and/or the biological information of interest (namely preventing any degradation thereof).       
 
         [0077]    Thus, said suspending solution may be a phosphate buffer (mono/dibasic, 10 mM EDTA, pH 8), a PBS buffer or a TE buffer (“Tris-EDTA”; 10 mM Tris, 10 mM EDTA (Ethylene Diamine Tetraacetic Acid), pH 8). Advantageously, said suspending solution can also comprise a preserving element which preserves the microbial fauna of the sample taken, namely that prevents any modification thereof, in order to allow the sample to be transported at ambient temperature from the site of sampling to the site of analysis, or else to defer the analysis step. A preservative of this type can, for example, consist of a preservative of “Cary-Blair” or “modified Cary-Blair” type, as described in the publication by Srijan et al. [1], or else a trypticase soy broth optionally supplemented with glycerol and/or 5% of sheep blood (in accordance with the teaching of the publication by Wasfy et al. [2]). 
         [0078]    Obviously, those skilled in the art will be able to adjust both the nature and the volume of the suspending solution according to the nature and the volume of sample taken. 
         [0079]    Advantageously, said suspending solution comprises a calibrator of the plant DNA, plant-derived bacterium or heavy peptides type, etc., making it possible to monitor and control the efficiency of the suspending, filtration and lysis steps (outside the device). In addition, the use of said calibrator is particularly advantageous for validating and interpreting the results. Furthermore, the use of said calibrator directly with the suspending solution makes it possible to dispense with an additional pipetting step in the protocol. 
         [0080]    The suspending solution can also comprise a means for capturing non-targeted elements (such as inhibitors, capable of interfering with the subsequent analysis steps). By way of example, active carbon or polyvinyl pyrrolidone (PVPP) can be used as means for capturing non-targeted elements in order to improve the sensitivity of detection, by retaining elements capable of disrupting/inhibiting the subsequent analysis steps. 
         [0081]    Moreover, when it is desired to analyze biological information contained in the biological matrix, preferably in the self-reproducible biological matrix, the suspending solution as defined above can, according to one particular embodiment of the invention, also contain a lysis solution, in order to induce the lysis of said biological material and the release of the biological information sought within said sample-suspending solution. 
         [0082]    According to one particularly preferred embodiment of the invention, said filtration means is selected from:
       a gradient filter, and   a superimposition of at least two filters, preferably of two or three filters, the pore size of which decreases from the interior to the exterior of the device.       
 
         [0085]    According to one preferred embodiment, the device is suitable for containing one or more filters each having a surface area greater than 50 mm 2 , preferably of between 100 and 350 mm 2 , advantageously between 290 and 330 mm 2 , in order to limit the clogging of the filters while at the same time ensuring a maximum filtered volume. 
         [0086]    The term “gradient filter” (also called “complex filter”) is intended to mean a filter which has a plurality of different pore sizes. More specifically, when a gradient filter is used for the purposes of the present invention, the size of the pores of this filter decreases from the interior to the exterior of the device in order to allow gradual and differential filtration of the sample with a heterogeneous matrix of interest. 
         [0087]    In any event, whether a gradient filter or a superimposition of at least two filters is used, as previously described, the pore size decreases from the interior to the exterior of the device, preferably from 500 μm to 0.1 μm, advantageously from 200 μm to 10 μm, particularly preferably from 200 μm to 20 μm. Quite obviously, the size of the pores of the filter(s) used is adjusted according to the size of the biological material and/or of the biological information that it is desired to recover in the filtrate. 
         [0088]    Thus, when it is desired to isolate, from the sample taken, viruses, toxins, metabolites, or other biological information present within said sample, a pore size of between 0.1 μm and 20 μm, preferentially 0.1 μm and 5 μm, can be used. 
         [0089]    In the case of a superimposition of at least two filters, a superimposition of two to four filters is preferably used. Advantageously, a superimposition of two or three filters is used. 
         [0090]    In one particular embodiment, said filtration means comprises at least one means for capturing non-targeted elements (such as inhibitors, capable of interfering with the subsequent analysis steps). By way of illustration, said means for capturing non-targeted elements can consist of active carbon, or of a material having a particular affinity for the inhibitors that may be contained in the biological material (proteins, polysaccharides, etc.), for example PVPP. 
         [0091]    According to one preferred embodiment, the container comprises at least one wall comprising at least one zone made of flexible material, suitable for undergoing a compression and generating, in response, an overpressure inside said container in order to allow or facilitate the filtration of the content through said filtration means. 
         [0092]    Said at least one zone made of flexible material proves to be particularly advantageous, insofar as the operator can generate, on said at least one zone, a manual pressure without excessive force, in order to allow/facilitate the filtration of the content through said filtration means, as previously indicated. The device according to the invention thus allows any laboratory technician, even the least experienced, to obtain, by using the device according to the invention, a filtrate containing the biological material and/or the biological information to be analyzed. Generally, the device according to the invention can fit to any system/device which makes it possible to facilitate the filtration operation. For example, said device can be fitted to a vacuum filtration system or to a centrifugation filtration system. It results from the aforementioned that the device according to the invention can, contrary to the protocol for obtaining biological material and/or biological information of the prior art, be easily automated, for example by means of an automated filtration device (or automatic filtration device), which is also a subject of the present invention. Such an automated filtration device, suitable for allowing the automation of the step of filtration of the content (namely of the suspension containing the sample with the heterogeneous matrix of interest), comprises:
       at least one site (and preferably a plurality of sites) suitable for receiving and maintaining the device for obtaining biological material and/or biological information according to the invention, preferably in the “inverted” or “upside down” position (namely in which the abovementioned opening essentially points downward),   a pressure member (for example a press such as a piston), which is mobile from an initial position (“resting position”) to a “pressure” position, in which said pressure member is suitable for exerting a pressure against said at least one zone made of flexible material, thereby having the effect of generating, in response, an overpressure inside said container, and thus allows the filtration of the content through said filtration means to a collecting receptacle (for example a tube, such as an Eppendorf® tube).       
 
         [0095]    Optimally, the automated filtration device according to the invention comprises up to eight or even ten different sites, thereby making it possible to simultaneously filter up to eight or even ten devices for obtaining biological material and/or biological information according to the invention. 
         [0096]    Preferably, the pressure member is actuated by a motor on command from the user or from an automated control center. For example, the user starts the motor by activating a switch (for example by pressing on a push-button), which has the effect of causing the pressure member to go from an initial position (“resting position”) to a “pressure” position, in which said pressure member exerts a pressure against said at least one zone made of flexible material. 
         [0097]    According to one particularly advantageous embodiment of the invention, the abovementioned automated filtration device is provided with an optical level detector (also called “optical barrier”) which stops the filtration step when the desired level of filtrate is reached within the collecting receptacle. More specifically, this optical detector operates by causing the pressure member to go from the pressure position to the resting position (under the action of the motor or, more simply, by stopping said motor), namely stops the pressure exerted by the pressure member on the zone made of flexible material when the optical level detector detects, via an optical sensor, that the desired level of filtrate is reached within the collecting receptacle, thus ending the filtration step. Without being bound by any theory, the fact that the pressure member ceases to exert a pressure against said at least one zone made of flexible material has the effect of ending the overpressure previously generated inside the container, which causes the filtration step to be stopped when the desired level of filtrate is reached within the collecting receptacle. An automated filtration device according to the invention, provided with an optical level detector, proves to be particularly advantageous, insofar as it makes it possible to collect the same volume of filtrate, this being whatever the type of stools, ensuring that the process for obtaining biological material and/or biological information using the device for obtaining biological material and/or biological information according to the invention is repeatable and robust. This is especially true given that, as previously explained, the calibrated hollow part of the device for obtaining biological material and/or biological information according to the invention makes it possible to take a given/predefined mass of sample with a heterogeneous matrix (without having to perform weighing operations), said mass being constant or virtually constant at each operation of taking a sample with a heterogeneous matrix. In other words, said calibrated hollow part and the automated filtration device according to the invention, provided with an optical level detector, act in synergy in order to guarantee optimal repeatability and robustness of the process for obtaining biological material and/or biological information using the device for obtaining biological material and/or biological information according to the invention. 
         [0098]    In one preferred embodiment, said stopper comprises at least one rigid zone, said rigid zone having a shape suitable for cooperating with at least one maintaining member, such as a clip, connected to a mixing apparatus, such as a vortex, so as to allow said device to be maintained on the mixing apparatus during the mixing of said device in order to facilitate the suspending of said sample in said suspending solution. 
         [0099]    According to one preferred embodiment, said rigid zone has a shape suitable for cooperating with a clip connected to said mixing apparatus, said rigid zone comprising:
       at least one anti-rotation means, comprising two distinct bearing surfaces, for example made up of two shoulders or of two tabs, each of the two distinct bearing surfaces being suitable for butting against or coming into abutment against one of the two ends of the clip when the stopper undergoes a rotational movement, in order to prevent or stop said rotation, and/or   at least one anti-translation means, comprising at least one bearing surface, such as a lip, a collar or a shoulder, said at least one bearing surface being suitable for butting against or coming into abutment against said clip when the stopper undergoes a translational movement, in order to prevent or stop said translational movement.       
 
         [0102]    Said at least one rigid zone allows the device of the invention to easily fit to the commercially available mixing/homogenizing apparatuses, such as apparatuses of vortex type, and to obtain results that are at the very least equivalent to those observed with devices specifically designed for this purpose, of the bead beater type. 
         [0103]    The abovementioned anti-rotation means and/or anti-translation means make(s) it possible to improve the maintaining of the device according to the invention by the maintaining member (for example a clip of the mixing apparatus of vortex type). 
         [0104]    The anti-rotation means makes it possible to limit, and preferably to eliminate, any rotation of said stopper (and thus, by extension, of the device according to the invention) relative to the abovementioned maintaining member. In addition to the fact that the anti-rotation means makes it possible to ensure entirely satisfactory maintaining between the stopper—and thus the device according to the invention—and the maintaining member of the mixing apparatus, this anti-rotation means also has the function of orienting the stopper, and consequently the calibrated sampling means, which is rigidly connected to said stopper, such that the calibrated hollow part of said sampling means is oriented downward or upward, preferably downward, when the device according to the invention is positioned on the maintaining member of the mixing apparatus. 
         [0105]    Regarding said anti-translation means, it makes it possible to limit, and preferably to eliminate, any translational movement of the stopper (and thus, by extension, of the device according to the invention) according to a vector that is oriented in the stopper-containing direction during the mixing operation. This anti-translation means thus makes it possible to ensure efficient mixing and to prevent the device according to the invention from getting away from the maintaining member during said mixing operation. Advantageously, this anti-translation means comprises at least one lip (protruding part) positioned on all or part of the surround of the upper part of the stopper. 
         [0106]    According to one particular embodiment, this anti-translation means consists of a collar (circular part) positioned on the upper part of the stopper and connected to the latter, having a diameter greater than the diameter of the body of the stopper. 
         [0107]    The lip formed on all or part of the surround of the upper part of the stopper constitutes a bearing face, which will cooperate with the abovementioned maintaining member (such as a clip) in order to guarantee the maintaining of the stopper—and thus of the device according to the invention—during the mixing operation. According to one preferred embodiment, when the maintaining member is a clip, the bearing face made up of the lip or the collar, as mentioned above, will come into abutment against said clip in order to prevent or stop any translational movement as explained hereinafter, under the title “detailed description of the invention”. 
         [0108]    According to one preferred embodiment of the invention, the upper part of the container—advantageously the shoulder of the flask should said container be a flask—performs the function of anti-translation means while making it possible to limit, or even eliminate, any translational movement of the stopper (and thus, by extension, of the device according to the invention) according to a vector that is oriented in the container-stopper direction during the mixing operation. Indeed, the bearing face made up of the upper part of said container (for example the shoulder of the flask) butts against/comes into abutment against said maintaining member in order to prevent or stop any translational movement of the device according to the invention according to a vector that is oriented in the container-stopper direction. 
         [0109]    According to one variant of the invention, a second anti-translation means can be positioned on all or part of the surround of the lower part of the stopper, in order to limit, or even eliminate, any translational movement of the stopper (and thus, by extension, of the device according to the invention) according to a vector that is oriented in the container-stopper direction during the mixing operation. 
         [0110]    Generally, said device is a device for obtaining biological material, preferably microbiological material, and said filtration means is suitable for allowing the selective passage of said biological material, preferably of said microbiological material, to the exterior of said device. 
         [0111]    A subject of the invention is also the stopper per se as previously defined. 
         [0112]    In addition, the invention also relates to the use of the abovementioned device for obtaining biological material and/or biological information from a sample with a heterogeneous matrix, preferably for obtaining biological material, preferably for obtaining microscopic biological material, advantageously for obtaining microbiological material. Said filtration means is suitable accordingly, in order to allow the selective passage of said biological material, preferably of said microscopic biological material, advantageously of said microbiological material, to the exterior of said device. 
         [0113]    According to one preferred embodiment, and as previously indicated, the sample with a heterogeneous matrix is selected from a soil sample, a stool sample, a sample of necrosed tissue, a food sample and an industrial sample, said sample preferably being a human or animal stool sample. 
         [0114]    Advantageously, said sample with a heterogeneous matrix is a stool sample. 
         [0115]    Another subject of the invention relates to a process for obtaining biological material and/or biological information from a sample with a heterogeneous matrix, said process using the device according to said process comprising the following steps:
       a) taking a predetermined volume of sample corresponding to a given mass using said calibrated sampling means,   b) suspending said sample in said suspending solution, optionally by mechanical suspending,   c) filtering the suspension obtained in step b) through said filtration means in order to obtain a filtrate containing said biological material and/or said biological information.       
 
         [0119]    According to one particular embodiment, the operator may store the suspension obtained following the abovementioned step b) and, where appropriate, defer the filtration step c). This makes it possible in particular to preserve the biological material and/or the biological information of interest for the purposes of subsequent analysis or analyses or, in the case of samples taken “in the field” (namely outside the laboratory), to send, to a laboratory, the device according to the invention comprising the suspension obtained in step b), namely comprising the biological material and/or the biological information of interest, obtained from the taking of a sample with a heterogeneous matrix. This in fact proves to be particularly advantageous when the samples with a heterogeneous matrix are taken in remote areas (in areas of the bush, for example), that are a long way from any analytical laboratory. In this embodiment, at least one preserving means suitable for the nature of the biological material and/or of the biological information to be analyzed is preferably used. Thus, said suspending solution may, for example, contain one or more chemical preservative(s) (for example of the Cary-Blair type) and/or one or more freezing mean(s) (allowing, by way of illustration, freezing at a temperature of about −80° C.). Quite obviously, the indispensable condition is that the preserving mean(s) used does (do) not (or, where appropriate, not significantly) modify the biological material and/or the biological information to be analyzed. 
         [0120]    According to one particular embodiment, said suspending solution comprises at least one culture means. This culture means may be at least one nutritive element or a plurality of nutritive elements (for example a culture medium) making it possible to promote the growth of all of the microorganisms present in the sample taken or to more or less selectively direct the growth of one or more type(s) of microorganisms. 
         [0121]    Said at least one culture means may also be at least one selective agent having inhibitory properties with regard to the growth of at least one type of microorganism, such as an antibiotic (bactericidal or bacteriostatic, preferably bactericidal) or an antifungal. The use of at least one selective agent can make it possible to direct the growth of at least one type of microorganism. 
         [0122]    Quite obviously, those skilled in the art will adjust the culture mean(s) present in the suspending solution according to the type(s) of microorganisms of interest. 
         [0123]    Another subject of the invention relates to a process for extracting biological information (for example nucleic acids, such as DNA and/or RNA) from a sample with a heterogeneous matrix, said process comprising the following steps:
       a) carrying out the abovementioned process for obtaining biological material and/or information in order to obtain a filtrate containing said biological material and/or said biological information,   b) when the biological information that must be extracted is contained within said biological material, such as a cell, a bacterium, a fungus or a yeast, carrying out a step of lysis of said biological material, preferably a step of mechanical lysis, in order to obtain a lysate comprising the biological information that must be extracted,   c) extracting the biological information from the filtrate obtained in step a) or from the lysate obtained in step b) by carrying out a suitable process for extracting biological information.       
 
         [0127]    According to one preferred embodiment, this extraction process is a process for extracting nucleic acid(s) (genetic information). In this embodiment, the abovementioned lysis step b) of the extraction process can be integrated into the extraction step c). For example, some automated systems for extracting genetic information of easyMAG® type (bioMérieux) integrate a lysis step into the extraction protocol. Said lysis may in particular be of chemical, mechanical or thermal type, depending on the nature of the sample with a heterogeneous matrix, on the biological material and/or biological information sought or else on the analytical techniques that will be subsequently used. 
         [0128]    According to one preferred embodiment, said process for extracting biological information comprises, before step c), and before step b) when said step must be carried out, a step of concentrating said biological material and/or said biological information, preferably by centrifugation (with or without density cushion of cesium chloride type for example) or by flocculation. When said concentrating step is carried out by centrifugation, the latter is carried out at a speed of between 6000 g and 12 000 g, advantageously of approximately 12 000 g. 
         [0129]    Another subject of the invention relates to a process for analyzing biological information, said process comprising the following steps:
       a) extracting said biological information to be analyzed using the abovementioned process for extracting biological information,   b) identifying and/or quantifying said biological information by any appropriate method of analysis, for example a method of genetic analysis such as a PCR, or a method of immunoassay or enzymatic assay type.       
 
         [0132]    According to one preferred embodiment, this analysis process is a process of genetic analysis (analysis of nucleic acid(s)). In this situation, said biological information is genetic information and the identification and/or quantification of this genetic information is (are) carried out by any appropriate method of genetic analysis, for example by PCR. 
         [0133]    In addition, a purification step may, as required, be carried out before step d), in order to remove any non-targeted element (such as one or more inhibitor(s) capable of interfering with the method(s) of analysis used in step d)). 
         [0134]    Another subject of the invention relates to a process for analyzing biological material from a sample with a heterogeneous matrix, said process comprising the following steps:
       a) obtaining a filtrate containing the biological material to be analyzed by using the abovementioned process for obtaining biological material,   b) where appropriate, inoculating a reaction medium with the filtrate obtained in step a), said reaction medium being suitable for allowing the growth and/or the expression of at least one metabolism of said biological material,   c) where appropriate, incubating the filtrate obtained in step a), or the inoculated reaction medium obtained in step b), for an appropriate period of time and at an appropriate temperature,   d) analyzing the biological material resulting from step a), b) or c) by any appropriate means of biological analysis.       
 
         [0139]    According to one preferred embodiment, said biological material is a microbiological material. 
         [0140]    In one particular embodiment, step a) of the process for analyzing biological material can be suitable for the analysis of “fragile” biological materials, such as Gram-bacteria, in particular using suspending means which cause fewer mechanical stresses on the bacteria walls. In order to allow suspending that is less damaging to these bacteria, the operator can reduce the mixing frequency, using mechanical suspending means which have a greater size, or else eliminate the use of mechanical suspending means. In the case where a mechanical suspending means would not be used, or where the sample would be difficult to suspend, a calibrated hollow part having at least one opening—preferably a plurality of openings—can be used in order to facilitate the suspending. Obviously, those skilled in the art will be able to adjust these parameters according to the biological material of interest termed “sensitive” and to the nature of the sample. 
         [0141]    Preferentially, the abovementioned process for analyzing microbiological material comprises step b) and optionally step c), advantageously steps b) and c), in which:
       step c), when it is present, consists in incubating the inoculated reaction medium obtained in step b) for an appropriate period of time and at an appropriate temperature, and   step d) of analyzing biological material consists of the detection and/or identification and/or counting of said biological material on/in said reaction medium, preferably by visual or optical reading.       
 
         [0144]    Another subject of the invention relates to a kit for obtaining biological material and/or biological information from a sample with a heterogeneous matrix, said kit comprising:
       said container and said stopper, in assembled or disassembled form,   at least one suspending solution, suitable for allowing said sample to be suspended,   preferably at least one mechanical suspending means, such as at least one bead.       
 
         [0148]    Advantageously, said container and said stopper are in disassembled form and are preferably packaged sterily. 
         [0149]    Another subject of the invention relates to the use of the abovementioned kit for carrying out said process for obtaining biological material and/or biological information. 
         [0150]    According to one preferred mode of the present invention, the device according to the invention is suitable for taking a sample of fecal material for the purposes of the microbiological analysis thereof. 
         [0151]    The biological material and/or biological information present in the filtrate obtained using the device according to the invention can be analyzed by various methods of biological analysis, such as enzymatic or immunological assays, nucleic sequence amplifications or tests on a Petri dish, or else by sequencing. In addition, the biological material and/or biological information present in the abovementioned filtrate can, where appropriate, be concentrated, purified, lysed or cultured, or enriched or be directly analyzed. 
         [0152]    By way of example, the biological material and/or biological information present in the abovementioned filtrate can be directly analyzed by MALDI-TOF mass spectrometry (depositing of at least one drop of filtrate on a MALDI-TOF plate), by Raman spectroscopy, by immunochromatography on a membrane or strip (lateral flow test), or by systems of bacteria identification of the API strip type. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0153]    The invention, its functionality, its applications and also its advantages will be more clearly understood on reading the present description, given with reference to the figures, in which: 
           [0154]      FIGS. 1A and 1B  show, respectively, a face-on view and a sectional view of a first embodiment of the device according to the present invention, 
           [0155]      FIG. 2  represents a face-on view of the stopper, separated from the container as represented in  FIGS. 1A and 1B , 
           [0156]      FIG. 3  is a view from below of the stopper of  FIG. 2 , 
           [0157]      FIG. 4  is a face-on view of a stopper according to a second embodiment, separated from the container according to the invention, 
           [0158]      FIG. 5  represents a view from below of the stopper represented in  FIG. 4 , 
           [0159]      FIG. 6  represents a side view of a device according to the invention connected to a mixing apparatus by means of a clip, 
           [0160]      FIG. 7  is a face-on view of the device according to the invention-clip-mixing apparatus assembly as represented in  FIG. 6 , 
           [0161]      FIGS. 8A and 8B  are perspective views of a stopper according to a third embodiment, 
           [0162]      FIG. 9  shows a face-on view of the pouring spout of the stopper represented in  FIGS. 8A  and  8 B, 
           [0163]      FIG. 10  is a perspective view of the upper plane of the central part of the stopper represented in  FIGS. 8A and 8B , 
           [0164]      FIG. 11  represents a perspective view of the lower plane of the central part of the stopper represented in  FIGS. 8A and 8B , 
           [0165]      FIG. 12  represents a face-on view of a second embodiment of the pouring spout of the stopper represented in  FIGS. 8A and 8B , 
           [0166]      FIGS. 13A and 13B  represent two perspective views of the sampling means suitable for being connected to the stopper represented in  FIGS. 8A and 8B , 
           [0167]      FIG. 14  represents a perspective view of a second embodiment of the calibrated sampling means according to the invention, 
           [0168]      FIG. 15  shows a perspective view of a third embodiment of the calibrated sampling means according to the invention, 
           [0169]      FIG. 16  is a perspective view of a stopper comprising a fourth embodiment of the calibrated sampling means according to the invention, 
           [0170]      FIGS. 17 and 18  represent, respectively, a face-on view and a side view of a fifth embodiment of the calibrated sampling means according to the invention, comprising a sliding extension in the “exit” position, namely conferring on the rod a first length of rod, as previously described, 
           [0171]      FIGS. 19 and 20  represent, respectively, a face-on view and a side view of the abovementioned fifth embodiment of the calibrated sampling means according to the invention, but in which the sliding extension is in the “re-entry” position, namely conferring on the rod a second length of rod, as previously described, less than the abovementioned first length of rod, 
           [0172]      FIG. 21  is a sectional view of an automated filtration device according to the invention, suitable for allowing the automation of the step of filtration of the content (namely of the suspension containing the sample with a heterogeneous matrix of interest) from the device for obtaining biological material and/or biological information according to the invention, 
           [0173]      FIG. 22  is a diagram representing schematically the various steps of the processes for analyzing biological information and biological material according to the present invention, 
           [0174]      FIG. 23  is a graphic representation of the results obtained for a linearity test, quantifying the DNA (by PCR, Cq number) of Gram negative bacteria (KPC) by means of the process for analyzing biological information according to the invention, as a function of the log of the concentration of KPC inoculated. A linear regression (R 2 ) is represented and calculated (Linear), 
           [0175]      FIG. 24  is a graphic representation of the results obtained for a linearity test, quantifying the DNA (by PCR, Cq number) of Gram positive bacteria (MRSA) by means of the process for analyzing biological information according to the invention, as a function of the log of the concentration of MRSA inoculated. A linear regression (R 2 ) is represented and calculated (Linear), 
           [0176]      FIG. 25  is a graphic representation of the results obtained for a linearity test, quantifying the DNA (by PCR, Cq number) of  S. pombe  yeasts by means of the process for analyzing biological information according to the invention, as a function of the log of the concentration of the  S. pombe  inoculated. A linear regression (R 2 ) is represented and calculated (Linear), 
           [0177]      FIG. 26  compares the quantification by PCR of adenovirus DNA using the Adenovirus R-Gene® kit (reference: 69-010B) with the QIAamp® DNA stool extraction kit (QiaGen) and the process for extracting biological information according to the invention, 
           [0178]      FIG. 27  (on the left, scale in ng/μl, on the right in ratio) presents the impact of the Bristol type of the stools on the amount and the purity of the microorganism DNA extracted. 
       
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
       [0179]    The objective of the detailed description hereinafter is to set out the invention sufficiently clearly and completely, in particular with reference to the abovementioned figures, but should not in any way be regarded as limiting the scope of the protection to the particular embodiments which are the subject of said figures. 
         [0180]    As represented in  FIG. 1A , the device according to the invention comprises a container  10 , which is in the form of a flexible plastic flask, that can be deformed in particular via a pressure exerted on said container by the operator&#39;s fingers. 
         [0181]    The flexible wall  101  of the abovementioned container  10  is made of flexible material. This flexible wall  101  contains, for example, rigid paper, cardboard, polyethylene, polyvinyl chloride, polypropylene, low-density polyethylene, polyethylene terephthalate, and also any suitable combination of these materials and/or of any other biobased material so that the flexible wall  101  of the container  10  has satisfactory properties in particular in terms of rigidity (in order to be able to place the container on a flat surface), of leaktightness and of deformation by compression when a pressure is exerted (for example by the operator&#39;s fingers or by the pressure member of an automated filtration device, as described above or below) on the flexible wall  101  of the container  10 , during the filtration step. 
         [0182]    As represented in this  FIG. 1A , the container  10  is closed by means of a stopper  11  comprising, from its upper part to its lower part:
       a tubular-shaped orifice  111 , making it possible in fine to harvest the filtrate (that can be closed by a cap  12 ),   a stopper body  112  (central part),   a calibrated sampling means  113  connected to the lower part of the stopper  11  (not numbered in  FIG. 1A ).       
 
         [0186]    More specifically, this calibrated sampling means  113  comprises a calibrated hollow part  1132  connected to the lower part of the stopper (not numbered on  FIG. 1A ) by means of a rod  1131 . The volume of the calibrated hollow part  1132  has been calculated beforehand to correspond to a predetermined mass of sample with a heterogeneous matrix, capable of containing the biological material and/or the biological information of interest. 
         [0187]    The device  1  is formed by assembly of the stopper  11  and of the container  10 . 
         [0188]    The container  10  is filled with a sufficient volume of suspending solution  13 , as previously defined. The level of suspending solution  13  is represented in  FIGS. 1A and 1B  purely by way of indication. 
         [0189]    The internal structure of the stopper  11 —and in particular of the stopper body  112 —is clearly visible in  FIG. 1B , which represents a sectional view of the device  1 . The stopper body  112  comprises an internal part  117  and an external part  119 , these two parts  117  and  119  delimiting a cavity within which a filtration means  116  is placed. This filtration means  116  consists of a superimposition of three filters  1161 ,  1162  and  1163 , each having a surface area of 314 mm 2 ; each of the three filters  1161 ,  1162  and  1163  also having pores of degressive sizes from the internal part  117  to the external part  119  of the stopper body  112 . More specifically, the first filter  1161  has pores of which the size is between 100 μm and 200 μm, whereas the two “upper” filters  1162  and  1163  each have pores of less than 50 μm in size. The first filter  1161  makes it possible to remove the coarsest entities (for example cell debris, etc.), whereas the two “upper” filters  1162  and  1163  make it possible to provide the selectivity of the filtration operation. As illustrated in  FIG. 1B , the filter  1163  having the smallest pore sizes (equal to those of the filter  1162 ) comes into contact with the external part  119  of the stopper body  112 . 
         [0190]    The rod  1131  of the calibrated sampling means  113  is connected to the internal part  117  of the stopper  11  by a connecting (or fixing) means  115 , placed on said internal part  117 . 
         [0191]    Openings  118  are made within the part  117  of the stopper  11  in order to allow the passage of the suspending solution  13  comprising the sample with a heterogeneous matrix (not represented) to the tubular-shaped orifice  111  via the filtration means  116 , comprising the superimposition of the three filters  1161 ,  1162  and  1163 . 
         [0192]    For the purposes of clarity, the use of the device  1  according to the invention is briefly described hereinafter. 
         [0193]    Using the hollow part  1132  of the calibrated sampling means  113 , the user samples a volume corresponding to the volume defined by the calibrated hollow part  1132  and corresponding to a desired mass of sample with a heterogeneous matrix. The container  10  with a flexible wall  101  is filled with a sufficient volume of a suspending solution  13 . As illustrated in  FIGS. 1A and 1B , the container  10  is closed/blocked by the stopper  11  (for example the stopper  11  is screwed onto a thread placed on the neck of said container) and, optimally, the tubular-shaped orifice  111  is closed by the cap  12 . 
         [0194]    The device  1  is then stirred in order to allow the suspending of the sample with a heterogeneous matrix contained in the calibrated hollow part  1132  of the sampling means  113 . Once the suspending has been carried out, the operator removes, as appropriate, the cap  12  and “turns” the device  1  “upside-down” (said device  1  thus being in an “inverted” or “upside-down” position, namely in which the tubular-shaped orifice  111  points essentially downward). Owing to this turning upside-down operation, the suspending solution  13 , loaded with sample with a heterogeneous matrix, passes through the openings  118  made within the internal part  117  of the stopper  11 , passes successively through the three filters  1161 ,  1162  and  1163  of the filtration means and flows to the exterior, in the form of a filtrate, by the tubular-shaped orifice  111 . The filtrate is then collected in any type of appropriate receptacle, for example in an Eppendorf tube. 
         [0195]    Advantageously, and as previously indicated, the passing of the suspending solution  13 , loaded with sample with a heterogeneous matrix, through the filtration means  116  is facilitated/accelerated when the operator exerts a pressure on the flexible wall  101  of the container  10 , resulting in an overpressure within said container  10  and in particular at the level of the filtration means  116 . This operation of compression of the flexible wall  101  by the operator proves to be particularly advantageous for facilitating the filtration, given the smallness/narrowness of the pores of the two upper filters  1162  and  1163 . 
         [0196]    The view represented in  FIG. 2  makes it possible to visualize the stopper  11 , when said stopper is disassembled from the container  10 , for example by unscrewing said stopper  11 . In this  FIG. 2 , the tubular-shaped orifice  111 , which can be closed by means of the cap  12 , the stopper body  112  and the calibrated sampling means  113  are distinctly observed. As represented in  FIGS. 1A and 1B , the latter comprises a rod  1131  connected, via one of its ends, to the internal part of the stopper body  112  and comprising, at the level of its opposite end, a calibrated hollow part  1132 . 
         [0197]    As previously indicated,  FIG. 3  is a view from below of the stopper  11 . Clearly seen in this  FIG. 3  is the radial arrangement of the openings  118 , made within the internal part  117  of the stopper  11 , around the rod  1131  of the sampling means  113 . Although the number and arrangement of the openings  118  can vary as desired by those skilled in the art, the radial arrangement of the openings  118  as represented in  FIG. 3  allows a good distribution of the volume of the suspending solution  13  loaded with sample with a heterogeneous matrix to the filtration means (not represented in  FIG. 3 ) via said openings  118 . 
         [0198]      FIG. 4  illustrates a second embodiment of a stopper  21  according to the invention. The stopper body  212  is provided:
       with an anti-rotation means  220 , and   with an anti-translation means  221 , both suitable for cooperating with a maintaining member such as a clip (not represented), of a mixing apparatus, such as a vortex (also not represented in  FIG. 4 ).       
 
         [0201]    The anti-rotation means  220  is positioned on the exterior surface of the stopper body  21 . As represented in  FIG. 4 , the anti-rotation means  220  is formed by an overthickness at the surface of the body of the stopper  21 . This overthickness defines two shoulders  2201  and  2202  on the stopper body  212 . 
         [0202]    When the maintaining member of the mixing apparatus (not represented in  FIG. 4 ) is a clip, each of the two branches of this clip will come to position itself on either side of the anti-rotation means  220 . When the stopper  21  is given impetus to rotate during the mixing step, one of the two shoulders  2201  or  2202  (depending on the direction of rotation) will come into abutment on the end of one of the two branches of the clip, preventing or stopping, de facto, the rotation of said stopper  21 . Preferably, the stopper is positioned on the clip such that the shoulders  2201  and  2202  come into abutment against the ends of each of the two branches of the clip. 
         [0203]    This anti-rotation means proves to be particularly advantageous insofar at it makes it possible to impose a particular orientation on the calibrated sampling means  213  connected to the internal part (not represented in  FIG. 4 ) of the stopper  21 . Thus, the anti-rotation element  220  is positioned on the external surface of the stopper body  212  in such a way that the calibrated hollow part  2132  of the sampling means  213  is oriented downward when the device comprising the stopper  21  is placed on the maintaining member (for example a clip) of the mixing apparatus of vortex type. This “downward” orientation of the hollow part makes it possible to improve the suspending of the sample with a heterogeneous matrix that it contains. 
         [0204]    The anti-translation means  221 , as represented in  FIG. 4 , is formed by a collar with a diameter greater than that of the stopper body  212 . The function of this anti-translation means  221  will be described hereinafter, in relation to  FIG. 5 . 
         [0205]    The arrangement of the anti-rotation means  220  and the anti-translation means  221  relative to the stopper body  212 , and in particular to the internal part  217  of this stopper body  212 , is clearly illustrated in  FIG. 5 , which represents a view from below of the stopper  21  according to the second embodiment of the invention, said stopper  21  being disassembled from the container  10 . As is the case in  FIG. 3 , the radial arrangement of the openings  218  around the rod  2131  of the sampling means  213  is here again observed. 
         [0206]      FIG. 6  makes it possible to visualize the maintaining of the device  2  (comprising the stopper  21  according to a second embodiment and the container  10 ) on a mixing apparatus  71  via the cooperation of the stopper body  212  and of a clip  40 , connected to the mixing apparatus  71 . In this regard, it should be noted that the clip  40  can be an integral part of the mixing apparatus  71  or can be connected thereto by any appropriate means. More specifically, it is observed, in this figure, that the shoulder  2201  of the anti-rotation means  220  comes into abutment against the end  402  of the corresponding branch of the clip  40 . Quite obviously, the shoulder  2202  also comes into abutment against the end  403  of the corresponding branch of the clip  40 , even though this cannot be observed in  FIG. 6 . In other words, the anti-rotation means  220  “butts” against each of the two ends  402  and  403  of the clip  40 , such that the stopper  21 , and consequently the device  2 , cannot undergo any rotational movement, in one direction or the other. Entirely advantageously, it is noted, in this  FIG. 6 , that the anti-rotation means  220  has been positioned on the external surface of the stopper body  212  so as to orient the calibrated hollow part  2132  “downward” when the stopper  21  is maintained by the clip  40 . As previously indicated, the orientation of the calibrated hollow part  2132  downward makes it possible to ensure optimal suspending of the sample with a heterogeneous matrix. 
         [0207]    In addition, the anti-translation means  221  is also clearly visible in  FIG. 6 . As previously indicated, it is, in this embodiment, a collar with a diameter greater than that of the stopper body  212 . When a translational movement occurs according to a vector that is oriented in the stopper  21 —container  10  direction, the collar acts as a bearing face, which comes into abutment against the part  401  of the clip  40 , thus preventing or stopping this translational movement. 
         [0208]    As represented in  FIG. 6 , the container  10  is filled with a sufficient volume of suspending solution  13 , as previously defined. The level of this suspending solution  13  is represented purely by way of indication in  FIG. 6 . The tubular-shaped orifice  221  of the stopper  21  is also closed by the cap  12 , in order to prevent any untimely exit of the suspending solution  13  during the mixing step. 
         [0209]      FIG. 7  represents a face-on view of the stopper  21 , maintained by the clip  40 , itself connected to the mixing apparatus  71 . In this  FIG. 6 , the positioning of the clip  40  on either side of the anti-rotation means  220 , such that each of the two shoulders  2201  and  2202  come into abutment against the corresponding ends  402  and  403  of each of the two branches of the clip  40 , is clearly distinguished. 
         [0210]    This clip  40  is also positioned under the anti-translation means  221 . In the event of there being a translational movement (according to a vector that is oriented in the stopper  21 —container  10  direction), the collar constituting the anti-translation means  221  will come into abutment against the part  401  (not represented in  FIG. 7 ) of the clip  40  so as to prevent or stop any translational movement. 
         [0211]    A third embodiment of the stopper  31  is illustrated in  FIGS. 8A and 8B . The latter clearly represents this stopper  31 , which comprises, from the upper part to the lower part:
       a tubular-shaped orifice  311 ,   an upper part  319 ,   an anti-translation means  321 ,   a stopper body  312 ,   an anti-rotation means  3201 ,  3202 ,   a calibrated sampling means  313  comprising a rod  3131  and a hollow part  3132 .       
 
         [0218]    In addition, the tubular-shaped orifice  311  can be closed by means of the cap  326 . 
         [0219]    As is possible to note in the light of  FIGS. 8A and 8B , this hollow part  3132  is oriented in a manner opposite to the anti-rotation means  3201 ,  3202 . In this way, when the device comprising the stopper  31  and the container  10  is maintained by a clip connected to a mixing apparatus, the anti-rotation means  3201 ,  3202  is oriented upward, as illustrated in  FIG. 6 , whereas said calibrated hollow part  3132  is deliberately oriented downward, in order to promote the suspending of the sample with a heterogeneous matrix present in the calibrated hollow part  3132 . 
         [0220]    In addition, contrary to the stopper according to a second embodiment of the invention, represented in  FIGS. 4-7 , in which the anti-rotation means  3201 ,  3202  is formed by an overthickness of material (for example an overthickness of plastic), positioned on the width of a part of the stopper body  212 , the anti-translation means  3201 ,  3202  of the stopper  31  comprises, over the entire length of the stopper body  312 , two parallel tabs  3201 ,  3202 , present at the surface of the stopper body  312 . 
         [0221]    As for the shoulders  2201  and  2202  represented in  FIG. 5 , when the stopper  21  is given impetus to rotate during the mixing step, one of the two tabs  3201 ,  3202  (depending on the direction of rotation) comes into abutment on the end of one of the two branches of the clip, preventing or stopping, de facto, the rotation of said stopper  31 . 
         [0222]    Preferably, the stopper  31  is positioned on the clip such that the tabs  3201 ,  3202  come into abutment against the ends of each of the two branches of the clip. 
         [0223]    The various parts constituting the stopper  31 , previously presented with reference to  FIGS. 8A and 8B , with the exception of the calibrated sampling means, are described hereinafter. 
         [0224]    The pouring spout  31 ′ of this stopper  31  is represented in  FIG. 9 . In said figure, a removable cap  326  having a shape complementary to that of the tubular-shaped orifice  311 , is observed, from the upper part to the lower part. This cap  326  can in particular be of cylindrical shape or of truncated conical shape. 
         [0225]    The pouring spout  31 ′ also comprises the upper part of the stopper body  319  and two rings,  327  and  328 , placed parallel to one another about the upper part of the stopper  319 . The assembly formed by the two rings  327  and  328  represents a male elastic interlocking element (commonly denoted “male clip-fastening element”) suitable for cooperating with a female elastic interlocking element (commonly denoted “female clip-fastening element”). The latter is obtained by reaming of the central element  31 ″, represented in  FIG. 10  and described hereinafter. 
         [0226]      FIGS. 8A and 8B  also represent a means for consistent positioning in a flattened area  323  of the anti-translation means  321 , making it possible to position the stopper relative to the ends  402  and  403  of each of the two branches of the clip  40  and to thus prevent the anti-translation means  321  from coming into contact with the mixing apparatus  71 . 
         [0227]      FIG. 10  makes it possible to precisely observe the upper part of the central element  31 ″, comprising various filter supports  325 ,  330 , suitable for receiving the filtration means represented by the numerical reference  116  in  FIG. 1B . Said filter supports comprise several lugs  325  positioned in proximity to the openings  318 , said lugs  325  extending as far as the level of the peripheral filter support  330 , consisting of a collar. As previously indicated, the central element  31 ″ also has a female clip-fastening element (not represented in  FIG. 10 ), comprising two grooves hollowed out by reaming in the internal side wall  329  of the central element  31 ″; these two grooves (not represented) being suitable for receiving, respectively, the reams  327  and  328  of the male clip-fastening element of the pouring spout  31 ′, and thus allowing the elastic interlocking of the pouring spout  31 ′ in the central element  31 ″. 
         [0228]    In addition, an opening  315  passes right through the central part of the central element  31 ″. This opening  315  has a shape that is complementary to that of the male clip-fastening means  3133  of the sampling means  313 , as represented in  FIGS. 13A and 13B . More specifically, said opening  315  is trapezoidal in shape and is proportioned such that, when the male clip-fastening means  3133  of the sampling means  313  (cf.  FIGS. 13A and 13B  below) is plugged into the opening  315 , the sampling means  313  is:
       (i) immobilized, and also   (ii) oriented relative to the anti-rotation means  3201 ,  3202 , as explained previously, in order to impose the desired orientation on the calibrated hollow part  3132  (cf. above).       
 
         [0231]    Furthermore, the complementarity between, on the one hand, the shape of the opening  315  and, on the other hand, that of the male clip-fastening means  3133  imposes and guarantees the desired orientation of the sampling means  313 —and in particular of its hollow part  3132 —during the operation of assembling the sampling means  313  with the central element  31 ″. In addition, the elastic interlocking of the part  3133  of the sampling means  313  in the opening  315  makes it possible to confer on the “central element  31 ”—sampling means  313 ″ assembly, sufficient rigidity so that the rod  3131  of the sampling means  313  does not curve during the taking of sample with a heterogeneous matrix, avoiding, by the same token, the risks of projection of sample with a heterogeneous matrix. 
         [0232]    The openings  318  as represented in  FIG. 10  allow the passage of the suspending solution  13  loaded with sample with a heterogeneous matrix of the container  10  to the intermediate part  340  of the central element  31 ″, containing the filtration means  116 . 
         [0233]    Although the assembly of the pouring spout  31 ′ with a central element  31 ″ and of the latter with the sampling means is obtained by elastic interlocking of male ( 327 ,  328  and  3133 ) and female (grooves hollowed out in the internal side wall  329  and opening  315 ) clip-fastening systems, the assembly of these various elements can, quite obviously, be obtained, alternatively, by any connecting/fixing system known to those skilled in the art, such as a screwing system, adhesive-bonding or else welding (for example heat-sealing). 
         [0234]      FIG. 11  represents a view from below, slightly in perspective, of the central part  31 ″ of the stopper  31 , on which the openings  318 , arranged radially around the opening  315 , are observed. By adjusting the relative placing of the trapezoidal opening  315  with respect to that of the anti-rotation means  3201 ,  3202 , the hollow part  3132  of the sampling means  313  is oriented such that said hollow part is oriented downward, in order to promote the suspending of the sample with a heterogeneous matrix (as previously explained). In this  FIG. 11 , it is also noted that the internal part  317  has a truncated cone shape, making it possible to limit the risks of leakage of the suspension comprising the sample with a heterogeneous matrix. During the screwing of the stopper onto the container, said truncated cone-shaped part comes into contact with the edge of the container and crushes the angle of the edge of the container over a small surface area, thus ensuring good leaktightness. 
         [0235]    Also observed in this  FIG. 11  is an internal thread  324  obtained by tapping, intended to cooperate with an external thread made on the upper part of the container  10 , preferably on the neck of the container  10 , in order to allow the assembly of the stopper  31  and of the container  10  by screwing. Quite obviously, other closure systems, preferably hermetic closure systems, may be used to allow the assembly of the stopper and of the container, for example an elastic interlocking (clip-fastening) system. 
         [0236]      FIG. 12  represents a second embodiment of the pouring spout  41 ′ of the stopper according to the invention. This pouring spout  41 ′ is suitable for being interlocked by elastic interlocking inside the central element  31 ″ of the stopper  31 . This is carried out by introducing the rings  427  and  428  into the corresponding grooves made in the internal side wall  329  of the central element  31 ″ (cf.  FIG. 10 ). 
         [0237]    As shown in this  FIG. 12 , the tubular-shaped orifice  411  of this pouring spout  41 ′ is rigidly connected to the breakable and repositionable cap  426  (for example, heat-sealed to said cap or simply molded at the same time as the cap, like conventional physiological saline pipettes), such that said breakable and repositionable cap  426  prevents, in this configuration, any uncontrolled exit of liquid via the tubular-shaped orifice  411 , to the exterior. 
         [0238]    In other words, when this breakable and repositionable cap  426  is rigidly connected to the tubular-shaped orifice  411 , this makes it possible to prevent any uncontrolled leakage of liquid to the exterior of the device according to the invention. Furthermore, the fact that this breakable and repositionable cap  426  is rigidly connected to the tubular-shaped orifice  411  and closes the latter indicates to the operate that no liquid has previously been poured by the pouring spout  41 ′. The breakable and repositionable cap  427  thus indirectly acts as an impregnability control. 
         [0239]    The breakable and repositionable cap  426  can be separated from the end of the tubular-shaped orifice  411  by the action of a force of tensile type or, preferably, twisting type on said breakable and repositionable cap  426 , in particular by exerting a rotational force on one, or preferably even two, of the wings  4261  and  4262  of said cap  426 . 
         [0240]    The end of the breakable and repositionable cap  426 , opposite the end in contact with the tubular-shaped orifice  411 , comprises a hollow part  4263  of substantially cylindrical or truncated conical shape. This hollow part  4263  has a shape complementary to that of the end of the tubular-shaped orifice  411 , such that, when the breakable and repositionable cap  426  is separated from the tubular-shaped orifice  411 , for example by applying a tensile force to one, or even two, of the wings  4261  and  4262  of said cap  426 , the latter can be used, subsequently, to again close the tubular-shaped orifice  411  of the pouring spout  41 ′. This closure is obtained by interlocking of the tubular-shaped orifice  411  in the hollow part  4263  of the breakable and repositionable cap  426 . 
         [0241]    It should be noted that, according to a simpler embodiment, the breakable and repositionable cap  426  can be replaced with a simple protective breakable part, which does not make it possible to reblock the tubular-shaped orifice  411  after separation of the protective breakable part thereof, contrary to the breakable and repositionable cap  426 , represented in  FIG. 12 . 
         [0242]      FIGS. 13A and 13B  represent, respectively, a perspective view of three quarters of the calibrated sampling means  313  and a back view of this same calibrated sampling means  313 . These two  FIGS. 13A and 13B  make it possible to observe the male clip-fastening means  3133 , the rod made of solid material  3131  and the oblong-shaped calibrated hollow part  3132  (having more specifically a “stick” shape), said calibrated hollow part  3132  having been obtained by hollowing out a region located at the end of the rod  3131  opposite the internal part of the stopper (not represented in  FIGS. 13A and 13B ). 
         [0243]    A second embodiment of the hollow part  4132  is represented diagrammatically in  FIG. 14 . On this face-on view, it is noted that said calibrated hollow part  4132  comprises openings  41321 , intended to facilitate the suspending of the sample with a heterogeneous matrix present at the level of the calibrated hollow part  4132 . This advantageous technical effect is obtained by allowing the suspending solution  13  to pass through the calibrated hollow part  4132  during the sample-suspending step. A calibrated hollow part  4132  according to this second embodiment proves to be particularly suitable for facilitating the suspending of samples with a heterogeneous matrix having significant adhesive properties, such as stools having a Bristol type 1 or 2. 
         [0244]      FIG. 15  illustrates a third embodiment of the calibrated hollow part  5132  having ovoid-shaped openings  51321  with dimensions greater than those of the openings  41321  represented diagrammatically in  FIG. 14 . These openings  51321  make it possible to further facilitate the suspending of a sample with a heterogeneous matrix contained within the calibrated hollow part  5132 . The hollow part  5132  according to this third embodiment proves to be particularly suitable for allowing the suspending of sample having adhesive properties that are more marked than those of the sample with a heterogeneous matrix that must be taken by means of or via the calibrated hollow part  4132  of  FIG. 14 , such as stools having a Bristol type 1-2. 
         [0245]      FIG. 16  represents one particular embodiment, in which the sampling means  613  comprises, on a large part of its rod  6131 , three square-shaped or rectangular-shaped calibrated hollow parts  6132 ,  6134  and  6135 , each having a predetermined volume so as to allow each of these three calibrated hollow parts  6132 ,  6134  and  6135  to sample a mass of 1000 mg. The calibrated sampling means  613 , represented in  FIG. 16 , thus makes it possible to take a total mass of 3000 mg of sample with heterogeneous matrix. A calibrated sampling means  613  of this type can prove to be particularly important for recovering, in fine, a filtrate containing a concentration of biological material and/or biological information that is sufficient to allow their analysis, in particular in the case where the biological material and/or biological information sought is (are) known to be present in very small amount in the material with a heterogeneous matrix from which the sample is taken. This sampling means  613  can also be used, advantageously, in veterinary applications, by increasing, as required, the volume defined by each of the hollow parts  6132 ,  6134  and  6135 . 
         [0246]    According to one particular embodiment, the rod  6131  of the sampling means  613  has, on its face opposite that comprising the three calibrated hollow parts  6132 ,  6134  and  6135 , three additional calibrated hollow parts. Preferably, these three additional calibrated hollow parts each make it possible to take a mass of sample with a heterogeneous matrix of approximately 1000 mg. Thus, this suitable sampling means comprising six calibrated hollow parts makes it possible to take a total mass of sample with a heterogeneous matrix of approximately 6000 mg of sample with a heterogeneous matrix. 
         [0247]    As represented in  FIGS. 17 to 20 , according to another preferred embodiment of the invention, the rod  7131  of the calibrated sampling means  713  is connected to a sliding extension  8 . This sliding extension  8  makes it possible to extend the length of the rod  7131  of said calibrated sampling means  713  to a first length of rod. This sliding extension  8  is positioned by a translation on the calibrated sampling means  713 , which already comprises a calibrated hollow part  7132  (which can be seen in  FIG. 20 ). When the sliding extension  8  is in “exit” position, namely making it possible to confer the abovementioned first length of rod (cf.  FIGS. 17 and 18 ), the operator can more easily take a sample with a heterogeneous matrix, for example a stool sample, at the bottom of a narrow tube, by means of at least one calibrated hollow part  81 , positioned at the level of the end of the sliding extension  8  furthest from the internal part of the stopper, while avoiding any contact between the stopper and the edges of said tube. 
         [0248]    After the sample with a heterogeneous matrix has been taken, and as previously indicated, the calibrated sampling means  713  and the container are assembled in order to make it possible to suspend the sample with a heterogeneous matrix and, in doing so, to continue the process for obtaining biological material and/or information according to the invention. During this assembly operation, the sliding extension  8  in the “exit” position (first length of rod; cf.  FIGS. 17 and 18 ) butts against the bottom of the container and, under the effect of the opposing resistance by the bottom of the container, the sliding extension  8  slides along the rod  7131 , in the direction of the internal part of the stopper, to its “re-entry” position (second length of rod; cf.  FIGS. 19 and 20 ), in order to allow the assembly of the stopper and of the container, for example by screwing the stopper onto a thread positioned on the neck of the container, as previously indicated. 
         [0249]    According to one embodiment of the invention, with a view to proceeding with the taking of sample with a heterogeneous matrix, the operator disassembles the stopper and the container, then slides the sliding extension  8  connected to the rod  7131  along the latter, from the “re-entry” position (second position; cf.  FIGS. 19 and 20 ) to the “exit” position (first position; cf.  FIGS. 17 and 18 ). According to one variant, in order to avoid any handling of the calibrated sampling means  713  prior to the sampling step (capable of generating microbial cross contamination), the stopper can be provided in a sterile packaging, the sliding extension  8  already in the “exit” position (first length of rod; cf.  FIGS. 17 and 18 ). Thus, the sample with a heterogeneous matrix can be directly taken without the operator needing to touch the sliding extension  8 . 
         [0250]    As previously indicated,  FIG. 21  is a sectional view of an automated filtration device  9  (or automatic filtration device) according to the invention. This automated filtration device makes it possible to automate the step of filtration of the content (namely of the suspension containing the sample with a heterogeneous matrix of interest) present within the device for obtaining biological material and/or biological information according to the invention. 
         [0251]    The operator positions the device for obtaining biological material and/or biological information according to the invention  1  in a site  91  suitable for receiving and maintaining the device  1  during the filtration operation. 
         [0252]    The operator then starts a motor  92  (for example by pressing on a push-button), which has the effect of causing a pressure member  93  to pass from an initial position (“resting position”—not represented in  FIG. 21 ) to a “pressure” position (visible in  FIG. 21 ), in which said pressure member  93  exerts a pressure against an area made of flexible material  101  of the container  10 . 
         [0253]    The pressure exerted by the pressure member  93  on said area made of flexible material  101  of the container  10  is maintained by virtue of the motor  92  for a period of time required to collect a desired volume of filtrate in a collecting receptacle  10000  (for example in an Eppendorf® tube). 
         [0254]    The automated filtration device  9  particularly advantageously comprises an optical level detector  94  (also called “optical barrier”). This optical level detector  94  makes it possible to stop the filtration step when the desired level of filtrate is reached in the collecting receptacle  10000 . More specifically, this optical detector operates by causing the pressure member  93  to pass from the pressure position to the resting position (under the action of the motor  92  or, more simply, by stopping the latter), namely stops the pressure exerted by the pressure member  93  on the area made of flexible material  101  of the container  10  when the optical level detector detects, via an optical sensor, that the desired level of filtrate is reached in the collecting receptacle  10000 , thus ending the filtration step. 
         [0255]    As previously explained, the fact that the pressure member  93  ceases to exert a pressure against said area made of flexible material  101  of the device  1  has the effect of ending the overpressure previously generated inside the container  10 , thereby resulting in the filtration step being stopped when the desired level of filtrate is reached in the collecting receptacle  10000 . 
         [0256]    As mentioned above, the automated filtration device  9 , provided with an optical level detector  94 , proves to be particularly advantageous insofar as it makes it possible to collect the same volume of filtrate, whatever the type of stools, giving the process for obtaining biological material and/or biological information using the device  1  according to the invention repeatability and robustness. This is especially true, as previously explained, given that the calibrated hollow part of the device  1  according to the invention makes it possible to take a given/predefined mass of sample with a heterogeneous matrix (without having to perform weighing operations), said mass being constant or virtually constant at each operation of taking a sample with a heterogeneous matrix. In other words, said calibrated hollow part and the automated filtration device  9  according to the invention, provided with an optical level detector  94 , act in synergy in order to guarantee optimal repeatability and robustness of the process for obtaining biological material and/or biological information using the device  1  according to the invention. 
         [0257]    Even though, in the interests of clarity, a single site  91  is represented on the automated filtration device  9  which is the subject of  FIG. 21 , this automated filtration device  9  optimally comprises up to eight or even ten different sites  91 , thereby making it possible to simultaneously filter up to eight or even ten devices  1 , this being with optimal repeatability and optimal robustness, as explained above. 
         [0258]      FIG. 22  is a diagram representing:
       (i) the various essential (solid line) and optional (dashed line) steps of the process for obtaining biological material and/or biological information according to the invention,   (ii) the various necessary (solid line) and optional (dashed line) steps of the various subsequent analysis pathways, allowing the identification and/or detection and/or quantification of said biological material and/or said biological information.       
 
         [0261]    Steps  171 ,  712  and  173 , represented on  FIG. 22 , are described in relation to the device  1 , represented in  FIGS. 1A and 1B . 
         [0262]    In order to carry out the sampling step  171 , the operator manually seizes the stopper, preferably by the stopper body, and fills the volume defined by the calibrated hollow part with the sample with a heterogeneous matrix (for example a stool sample). The stopper and the container, filled beforehand with a suspending solution, are assembled, for example by screwing the stopper onto the neck of the container, such that the sample with a heterogeneous matrix contained in the hollow part is in contact with the suspending solution. 
         [0263]    It should be noted that the container may contain, in addition to the suspending solution, one or more suspending means, for example 30 glass beads, each having a diameter of 3 mm. 
         [0264]    The suspending step  172  can be carried out by agitating the device in order to allow the suspending of the sample with a heterogeneous matrix contained in the calibrated hollow part in the suspending solution. In order to facilitate and/or accelerate this mechanical suspending step  172 , the device can be connected to a mixing means, such as a vortex, as represented in  FIG. 6 . Advantageously, the device is connected to said mixing means by a maintaining member which closely cooperates with the rigid part of the stopper, in order to ensure effective maintaining of the device during the mechanical suspending step  172 , consisting, in the case in point, of a mixing step. 
         [0265]    Once the operator considers that the sample with a heterogeneous matrix has been correctly suspended in the suspending solution, the suspending step  172  is finished. At this time, the device containing the suspension of sample with a heterogeneous matrix can optionally be stored, in step  1721 , preferably after addition of at least one preserving means and/or by freezing. Alternatively, the device can also be incubated at a temperature, for example at 37° C., for a period of a few hours to several days in order to enrich the suspension of certain microorganisms present in the sample. This incubation step can in particular be carried out in the presence of a selective culture means contained in the device according to the invention. The maximum duration of storage depends quite obviously on the preserving means used, where appropriate, and on the nature of the biological material and/or of the biological information to be analyzed. As previously indicated, this storage step can prove to be particularly advantageous in the context of operations for taking samples with a heterogeneous matrix “outside the laboratory”. Indeed, the device optionally comprising a storage means can, after the tubular-shaped orifice has been closed by a cap, be sent to an analytical laboratory for the purpose of carrying out all the desired analyses of the biological material and/or of the biological information that may be contained in the sample with a heterogeneous matrix taken in step  171 . 
         [0266]    Alternatively, the operator can directly carry out the filtration step  173 . In order to carry out this filtration step  173 , the removable closure means is removed from the tubular-shaped orifice, then the device is subsequently turned upside down and at least one pressure is applied to at least one area of the flexible wall of the container, in order to create an overpressure in this container and thus to force the suspending solution loaded with sample with a heterogeneous matrix to pass through the filtration means of the stopper, via the openings. Thus, the suspending solution loaded with sample with a heterogeneous matrix which flows along the tubular-shaped orifice has necessarily been filtered by the filtration means  116  positioned in the body of the stopper. At the end of this filtration step  173 , the filtrate containing said biological material and/or biological information is poured, via the tubular-shaped orifice, into a collecting tube, for example an Eppendorf tube. Once said biological material and/or biological information has (have) been isolated in the collecting tube, the operator can carry out various biological analyses, according to the nature of the biological material and/or biological information sought. 
         [0267]    When it is desired to analyze the viable biological material, conventional microbiology techniques  1751  could be carried out for these purposes, such as the inoculating and culturing of said viable biological material on solid, liquid or semi-solid, selective or non-selective reaction media, preferably comprising a culture medium. The metabolic expression of this viable biological material can also be evaluated via the use of enzymatic tests (for example API strips, MALDI-TOF plate, lateral flow test). These microbiology techniques make it possible to detect and/or identify and/or count, in step  1752 , the viable biological material sought. However, depending on the analysis techniques used, and on the type of viable biological material of interest, an additional step of enrichment may prove to be necessary in order to promote the growth of said biological material and thus to increase the concentration thereof in the reaction medium. 
         [0268]    Quite obviously, said viable biological material, just like the biological information in the broad sense, can also be the subject of genetic, protein and/or metabolic analyses. To this effect, the reader will refer to the analysis of biological information, which is described hereinafter. 
         [0269]    After the filtration step  173 , when the operator wishes to analyze the biological information contained in the filtrate, and comprising in particular metabolites, proteins or else nucleic acids, specific detection and/or identification and/or quantification techniques are carried out. 
         [0270]    Particularly advantageously, a step of concentrating said biological information  1741  contained in the filtrate is carried out by centrifugation at 6000-12 000 g of by flocculation, in order to concentrate the biological information of interest and to reduce the amount of non-targeted elements (such as inhibitors) present in the suspension. Where appropriate, a lysis step  1742  is carried out, when the biological information of interest is contained in said biological material, in particular when said biological material is a self-reproducible biological material, in order to make the biological information accessible to the analysis mean(s). 
         [0271]    According to one variant, the lysis step  1742  can be carried out before the concentrating step  1741 . 
         [0272]    The subsequent steps depend on the nature of the biological information sought and on the analysis techniques used, genetic, protein or metabolic analysis techniques. When the operator wishes to analyze the biological information of interest by means of a genetic analysis technique (analysis of nucleic acid(s)), a nucleic acid extraction step  17431  is carried out by implementing any suitable nucleic acid extraction protocol. The nucleic acids obtained at the end of the extraction step  17431  are detected and/or identified and/or quantified by any appropriate genetic analysis method  174132 , for example by PCR. 
         [0273]    The protein and/or metabolic analysis  17441  involves, for its part, suitable analysis techniques, such as immunological assays (immunoassays) or else enzymatic assays (non-limiting list), which make it possible, in step  17450 , to detect and/or identify and/or quantify the proteins and/or the metabolites of interest, initially present in the sample with a heterogeneous matrix. 
         [0274]    The examples hereinafter will make it possible to understand the present invention more clearly. However, these examples are given only by way of illustration and should in no way be regarded as limiting the scope of said invention in any way. 
       Examples 
     Example 1—Assembly of a Device According to the Invention 
       [0275]    For the purposes of example 1, the device for obtaining biological material and/or biological information from a sample with a heterogeneous matrix is assembled in the following way:
       inserting the first filter  1161  (Filtrona reference: BNW440148) into the central part of the central element  31 ″ of the stopper  31 ,   inserting a second filter  1162  (Pall Pad reference: 66025) in superimposition with respect to the first filter  1161 ,   putting in place the pouring spout  41 ′ by elastic interlocking of the male clip-fastening systems  427  and  428  and the female clip-fastening systems (grooves hollowed out in the internal side wall  329  of the central element  31 ″),   clip-fastening the calibrated sampling means  313 , having a calibrated hollow part  3132  (volume: 300 μl), in the opening  315  of the central element  31 ″,   adding 30 glass beads having a diameter of 3 mm to the container  10 ,   adding 5 ml of suspending solution (in the case in point suspending buffer of TE buffer type) to the container  10 , then   screwing the stopper comprising the pouring spout  41 ′ and the calibrated sampling means  313  onto the neck of the container  10 .       
 
       Example 2—Process for Obtaining Biological Material and/or Biological Information Using the Device of Example 1 
       [0283]    The protocol for obtaining biological material and/or biological information from a sample with a heterogeneous matrix according to the present invention, using the device described in example 1, comprises the following steps: 
         [0284]    1) Taking the Sample 
         [0285]    More specifically, the user firstly opens the pot containing the stools to be analyzed, and takes a sample using the calibrated sampling means  313 , such that the calibrated hollow part  3132  of the calibrated sampling means  313  is filled with the sample of interest. In order to allow optimal calibration, the calibrated sampling means  313  is then “scraped” on the lip of the abovementioned pot in order to remove the possible surplus of salts present on the calibrated sampling means  313  and/or overflowing the calibrated hollow part  3132 . 
         [0286]    2) Suspending the Sample 
         [0287]    Secondly, the user introduces the calibrated sampling means  313  into the container  10 , and screws closed the stopper  31  before attaching the device according to the invention onto a mixing apparatus  71  (Genie II vortex; Scientific Industries, Inc.). More specifically, as represented in  FIG. 6 , the stopper of the device according to the invention is maintained by means of a clip  40  connected to a “vortex platform”, itself placed on the mixing apparatus  71  (“vortex”). The device thus attached is mixed (“vortexed”) until complete suspension of the sample is obtained, namely for a period of time generally of between 30 seconds and approximately 2 minutes depending on the Bristol type. Quite obviously, the optimal mixing (“vortexing”) time for the sample may easily be determined by the user on the basis of their general knowledge and, where appropriate, of routine tests. 
         [0288]    3) Filtering the Sample Suspended in Step 2) 
         [0289]    In a third step, once the suspending step has been completed, the user removes the device according to the invention from the “vortex platform”, then removes the cap  426  by applying a tensile force to the wings  4261  and  4262 , turns the device upside down above a collecting tube (2 ml Eppendorf), then exerts a pressure with their fingers on the flexible walls  101  of the container  10  until the desired volume of filtrate is collected, namely approximately 2 ml. The filtrate thus obtained is ready to be analyzed. 
       Example 3—Efficiency of the Calibration of the Sampling Means  313  on Samples of Stools of Various Bristol Types 
       [0290]    The sampling is carried out using stool samples from healthy donors, stored at a temperature of −80° C. The stools are thawed beforehand. 
         [0291]    The samplings are carried out by implementing step 1 of the process of example 2. 
         [0292]    In order to accurately determine the mass of stools sampled in the calibrated hollow part  3132  of the sampling means  313 , the stopper is weighed before (tare) and after the sampling operation. 
         [0293]    The results obtained are presented in table 1 below: 
         [0000]    
       
         
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Bristol 
                 Bristol 2 
                 Bristol 3 
                 Bristol 3 
                 Bristol 4 
                 Bristol 5 
                 Bristol 6 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Weight of stools 
                 189 
                 201 
                 214 
                 217 
                 217 
                 191 
               
               
                 per sampling 
                 199 
                 200 
                 215 
                 271 
                 209 
                 191 
               
               
                 (mg) 
                 143 
                   
                 259 
                 223 
                   
                 211 
               
               
                   
                 206 
                   
                 242 
                 216 
                   
                 236 
               
               
                   
                   
                   
                 207 
                 187 
                   
                 220 
               
               
                   
                   
                   
                 216 
                 258 
                   
                 234 
               
               
                   
                   
                   
                 220 
                 236 
               
               
                   
                   
                   
                 254 
                 224 
               
               
                   
                   
                   
                 209 
                 204 
               
               
                   
                   
                   
                 192 
                 244 
               
               
                   
                   
                   
                 192 
                 191 
               
               
                   
                   
                   
                 198 
                 200 
               
               
                 Mean 
                 184 
                 201 
                 218 
                 223 
                 213 
                 214 
               
               
                 CV (%) 
                 15 
                 0.4 
                 10 
                 12 
                 3 
                 9 
               
               
                   
               
             
          
         
       
     
         [0294]    Example 3 makes it possible to calculate the sampling variability (CV), corresponding to all of the steps implemented in order to carry out the sampling. 
         [0295]    The CV is a percentage calculated with respect to the standard error of the mean, by applying the following formula: 
         [0000]    
       
         
           
             
               
                 ∑ 
                 
                   
                     ( 
                     
                       x 
                       - 
                       
                         x 
                         _ 
                       
                     
                     ) 
                   
                   2 
                 
               
               n 
             
           
         
       
     
         [0000]    where {tilde over (x)} the mean (number 1, number 2 . . . ) of the sample and n is the size of the sample. 
         [0296]    The calibrated hollow part  3132  of the sampling means  313  makes it possible to collect a given/predetermined mass of stools easily, efficiently and reproducibly (robustness of the process for obtaining biological material and/or biological information using the device according to the invention), this being for various Bristol types. The minor variations in mass observed by repeatability are considered to be acceptable in that they do not significantly affect the amount of biological material and/or of biological information obtained by using the device. 
       Example 4—Test of Linearity of Detection of the Nucleic Acids of Various Types of Microorganisms Using the Process for Analyzing Biological Information According to the Invention 
       [0297]    In order to verify the efficiency of the process for analyzing biological information (in the case in point DNA) from a sample with a heterogeneous matrix, which is the subject of the present invention, the following three types of microorganisms were inoculated into the suspending solution contained in the device according to the invention:
       carbapenem-resistant  Klebsiella pneumoniae  (KPC),   methicillin-resistant  Staphylococcus aureus  (MRSA),     Schizosaccharomyces pombe  ( S. pombe ).       
 
         [0301]    These three types of microorganisms were chosen according to their size and their characteristics, KPC being a Gram-negative  bacillus  5 μm long, resistant to carbapenems, MRSA being a Gram-positive coccus 1 μm in diameter, resistant to methicillin, and  S. pombe  being a yeast 10 μm long and cylindrical in shape. 
         [0302]    The inoculation of the suspending solution is prepared in the following way: 
         [0303]    —MRSA and KPC 
         [0304]    MRSA and KPC are cultured on TSA (trypticase soy agar) overnight at 37° C. A solution of 7 McFarland corresponding to 2×10 9  CFU/ml (use of a densitometer) is prepared and diluted in cascade (1/10 dilutions) until the dilution 10 3  CFU/ml is obtained. 100 μl of the dilutions concerned are used to inoculate the suspending solution before the addition of the stool sample. 100 μl of the dilution at 10 3  CFU/ml are then deposited on COS dishes (reference: 43041, bioMérieux) (Columbia Agar+5% sheep blood) in triplicate in order to verify the concentration of the inoculant. The colonies are counted after 24 h of incubation (37° C.) 
         [0305]    — S. pombe    
         [0306]      S. pombe  is cultured for two to three days on SDC agar (glucose Sabouraud agar; reference: 43555, bioMérieux) at 30° C., then overnight in a Sabouraud broth (30° C.) (reference: 42108, bioMérieux), the optical density is measured on a spectrophotometer and the broth is diluted in cascade until the dilution 10 5  CFU/ml is obtained. 100 μl of the dilutions concerned are used to inoculate the suspending solution before the addition of the stool sample. Direct counting of the yeasts under a microscope using a Kova cell is then carried out using the dilution at 10 6  CFU/ml in order to verify the concentration of the inocula. 
         [0307]    Each suspending solution of each device is respectively inoculated with the following amounts of MRSA and of KPC: 0, 10 5 , 10 7 , 10 8  and 2×10 9  CFU. For  S. pombe , the amounts inoculated are: 0, 10 5 , 10 6  and 10 7  CFU. 
         [0308]    Three controls are systematically carried out during the tests:
       Negative control no. 1: device according to the invention, comprising a non-inoculated suspending solution (TE buffer) and a stool sample analyzed beforehand in order to be sure of the absence of MRSA, of KPC and of  S. pombe  in these stools,   Negative control no. 2: device according to the invention comprising a non-inoculated suspending solution (TE buffer), without stool sample, in order to verify that there have not been any contaminations during the manipulations,   Positive control: device according to the invention comprising a suspending solution (TE buffer) inoculated with all of the three microorganisms tested, at a known concentration, without stool sample, in order to determine the impact of the stools on the quantitative PCR detection of the microorganisms.       
 
         [0312]    The following were added to the container  10 : 5 ml of TE buffer and 30 glass beads having a diameter of 3 mm. The inoculation of the suspending solution is then carried out by adding 100 μl of the previously prepared suspensions. After inoculation, the device is used to carry out the sampling, suspending and filtration steps as described in example 2. 
         [0313]    The sampling step 1) is carried out using a sample of stools from healthy donors, of stools stored in a pot and frozen at −80° C. The stools known to naturally contain KPC, MRSA or  S. pombe  were discarded. 
         [0314]    The filtrate collected at the end of the filtration step 3) (cf. example 2) is centrifuged for 3 minutes at 12 000 g in order to concentrate the microorganisms. The supernatant is discarded and the pellet is resuspended in 600 μl of TE buffer by vortexing vigorously. 
         [0315]    The 600 μl thus obtained are then transferred into a lysis tube (1.5 ml Eppendorf) prefilled with a mixture of beads consisting of 150 μg of zirconium beads 0.1 mm in diameter and 600 μg of glass beads 1 mm in diameter. The Eppendorf tube is subsequently placed on a “vortex platform” (24-position horizontal platform), itself placed on a Genie 2 vortex (Scientific Industries, Inc.), for 20 minutes and at maximum power, in order to lyse the microorganisms contained in the Eppendorf tube. 
         [0316]    The solution thus lysed is recovered by pipetting and the beads are washed with 200 μl of TE buffer. The TE buffer that was used to wash the beads is recovered and added to the solution previously recovered by pipetting. 
         [0317]    The entire volume recovered is then separated into two aliquots, each aliquot being introduced into a tube containing 2 ml of easyMAG® lysis buffer (reference: 280134, bioMérieux) and 140 μl of easyMAG® silica (reference: 280133, bioMérieux). Each mixture is then placed in a well of an easyMAG® shuttle (bioMérieux). Specific protocol “B” is launched, including an “off-board” lysis and an elution in a volume of 50 μl. The 2×50 μl of eluate originating from one and the same device are mixed, after extraction, in one and the same tube. 
         [0318]    The easyMAG® reagents and consumables are listed hereinafter: 
         [0000]    Reference: 280134 NucliSENS® easyMAG® lysis buffer (4×1000 ml/bottle)
 
Reference: 280130 NucliSENS® easyMAG® extraction buffer 1 (4×1000 ml/bottle)
 
Reference: 280131 NucliSENS® easyMAG® extraction buffer 2 (4×1000 ml/bottle)
 
Reference: 280132 NucliSENS® easyMAG® extraction buffer 3 (4×1000 ml/bottle)
 
Reference: 280133 NucliSENS® easyMAG® magnetic silica (48×0.6 ml/flask)
 
Reference: 280135 NucliSENS® easyMAG® consumables
 
Reference: 280146 Tips for multipette.
 
         [0319]    After extraction, 5 μl of eluates are used to carry out a PCR analysis targeting each of the three microorganisms inoculated. The results are expressed in Cq (quantification cycle), which is directly linked to the concentration of DNA present in the amplification tube as a function of the log of the concentration of microorganisms inoculated. 
         [0320]    The primers and the probes used for the molecular analysis are specific for the KPC genes (gene encoding carbapenemases) in  K. pneumoniae , for the SCCmec genes (methicillin resistance gene) in MRSA and for the SPBPJ4664.02 genes (glycoprotein) in  S. pombe.    
         [0321]    For each of the sequences of interest, the PCR mixes are prepared in accordance with the indications given in tables 2-4 below: 
         [0000]    
       
         
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 PCR reagent mix for analysis of KPC 
               
             
          
           
               
                   
                 Reagents 
                 Final concentration 
               
               
                   
                   
               
               
                   
                 Acros water 
                 — 
               
               
                   
                 Buffer pH 8.6 (X) 
                 1 
               
               
                   
                 MgCl 2  solution (mM) 
                 3.5 
               
               
                   
                 dNTP (mM) 
                 0.2 
               
               
                   
                 BSA (μg/μl) 
                 0.5 
               
               
                   
                 Antisense primer (μM) 
                 0.2 
               
               
                   
                 Sense primer (μm) 
                 0.2 
               
               
                   
                 Probe (μM) 
                 0.1 
               
               
                   
                 DNA polymerase (Fast Start) (U/μl) 
                 0.08 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 PCR reagent mix for analysis of  S. pombe   
               
             
          
           
               
                   
                 Reagents 
                 Final concentration 
               
               
                   
                   
               
               
                   
                 Acros water 
                 — 
               
               
                   
                 Buffer pH 8.6 (X) 
                 1 
               
               
                   
                 MgCl 2  solution (mM) 
                 5 
               
               
                   
                 dNTP (mM) 
                 0.2 
               
               
                   
                 BSA (μg/μl) 
                 0.5 
               
               
                   
                 Antisense primer (μM) 
                 0.65 
               
               
                   
                 Sense primer (μm) 
                 0.65 
               
               
                   
                 Probe (μM) 
                 0.17 
               
               
                   
                 DNA polymerase (Fast Start) (U/μl) 
                 0.08 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 PCR reagent mix for analysis of MRSA 
               
             
          
           
               
                   
                 Reagents 
                 Final concentration 
               
               
                   
                   
               
               
                   
                 Acros water 
                 — 
               
               
                   
                 Buffer pH 8.6 (X) 
                 1 
               
               
                   
                 MgCl 2  solution (mM) 
                 5 
               
               
                   
                 dNTP (mM) 
                 0.2 
               
               
                   
                 BSA (μg/μl) 
                 0.5 
               
               
                   
                 Antisense primer (μM) 
                 0.2 
               
               
                   
                 Sense primer (μm) 
                 0.2 
               
               
                   
                 Probe (μM) 
                 0.1 
               
               
                   
                 DNA polymerase (Fast Start) (U/μl) 
                 0.144 
               
               
                   
                   
               
             
          
         
       
     
         [0322]    For each PCR reaction, the PCR reagent mixes are prepared in a final volume of 20 μl, to which are added 5 μl of the eluates obtained for each sample treated. The mixes are then amplified using a thermocycler, according to the amplification cycles presented in table 5 below: 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
               
               
               
             
           
               
                   
                 TABLE 5 
               
               
                   
                   
               
               
                   
                 Enzymatic 
                   
                   
               
               
                   
                 activation 
               
               
                   
                 (Fast 
               
               
                   
                 Start) 
                 Denaturation 
                 Hybridization/elongation 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                 Temperature 
                 95 
                 95 
                 65° C. 
               
               
                 (° C.) 
               
               
                 Time 
                 5 min 
                 15 sec 
                 45 sec 
               
             
          
           
               
                 Cycle(s) 
                  1 
                 50 
               
               
                   
               
             
          
         
       
     
         [0323]    The results obtained (presented in  FIGS. 23, 24 and 25 ) show a linear relationship between the amount of microorganisms previously inoculated into the suspending solutions and the amount of PCR-amplified DNA found in the eluate after extraction. Consequently, this demonstrates that the process for analyzing biological information according to the invention makes it possible to detect, in a “LOG-linear” manner, the microorganisms present in the stool samples, this being over the entire range of amounts tested. 
         [0324]    In conclusion, in the light of the results thus obtained, it appears that the process for analyzing biological information according to the invention is efficient for isolating and identifying the microorganisms present in stool samples (including yeasts), both quantitatively and qualitatively. 
       Example 5—Comparison of the Process for Analyzing Biological Information According to the Invention and of the Protocol Using the QIAamp® DNA Stool Kit (QiaGen; Reference: 51504) 
       [0325]    The inocula were prepared according to the methodology set out in example 4, from a solution at 0.5 McF corresponding to 10 8  CFU/ml. The suspending solution is inoculated with 10 8  CFU of MRSA,  S. pombe  and KPC. The control of the inocula is carried out as in example 4. The same is true for the negative controls no. 1 and no. 2 and of the positive control. 
         [0326]    The obtaining and the quantification of the biological information are carried out by implementing the process for analyzing biological information (in the case in point DNA) according to the invention, described in example 4. 
         [0327]    The obtaining of biological information with the QIAamp® DNA stool kit is carried out according to the experimental protocol provided by the supplier (QiaGen). Nevertheless, considering the present metagenomic application, a mechanical lysis step was added after addition of the ASL buffer and before incubation at 95° C. for 10 minutes, in order to obtain a better microorganism lysis yield. This is part of the general knowledge of those skilled in the art. 
         [0328]    The biological information obtained using the QIAamp® DNA stool kit is quantified under the same conditions as those which make it possible to quantify the biological information obtained by implementing the process according to the present invention. 
         [0329]    The results obtained for each of the two protocols are presented in table 6 below. 
         [0000]    
       
         
               
             
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 Comparison of the results obtained by 
               
               
                 quantitative PCR using the DNA extracted by the process 
               
               
                 according to the invention and by the process according 
               
               
                 to the QIAamp ® DNA stool kit (QiaGen) 
               
             
          
           
               
                   
                 Amount of 
                   
               
               
                   
                 DNA 
               
               
                   
                 extracted 
                 qPCR result (Cq) 
               
             
          
           
               
                   
                 Protocol 
                 (μg) 
                 MRSA 
                 KPC 
                 
                   S. pombe 
                 
               
               
                   
                   
               
               
                   
                 Invention 
                 2.5-3 
                 26.6 
                 28.1 
                 35.2 
               
               
                   
                 QIAamp ® 
                   7-10 
                 28.4 
                 29.2 
                 Not 
               
               
                   
                 DNA stool 
                   
                   
                   
                 detected 
               
               
                   
                   
                   
                   
                   
                 (&gt;40) 
               
               
                   
                   
               
             
          
         
       
     
         [0330]    This table 6 presents the amount of DNA extracted for each of the two protocols, and also the value of the Cqs obtained by the PCR analysis (Cq corresponds to the Ct or threshold cycle, the cycle at which the emitted fluorescence value reaches the predefined threshold). 
         [0331]    The results thus obtained clearly demonstrate that the molecular analysis of the biological information (DNA) by PCR, carried out using the eluates obtained by implementing the process according to the invention is efficient and exhibits improved sensitivity compared with the process using the QIAamp® DNA stool kit. In addition, it is important to note that only the process according to the invention makes it possible to detect yeasts (in this case,  S. pombe  yeasts). 
         [0332]    In conclusion, the process for analyzing biological information according to the invention, using the device according to the invention, allows efficient detection of the biological information contained in the microorganisms present in a stool sample (including regarding yeasts), while at the same time simplifying the steps of sampling 1), suspending 2) and filtration 3) of the samples, as indicated in example 2. 
       Example 6—Comparison of the Process for Analyzing Biological Information According to the Invention and of the Process Using the QIAamp® DNA Stool Kit for a Human Organism Microbiota Sequencing Application 
       [0333]    The QIAamp® DNA stool kit (QiaGen; reference: 51504) is used in accordance with the experimental protocol provided by the supplier (QiaGen) and modified as mentioned in example 5 above. 
         [0334]    The devices which are the subject of the present invention are assembled in accordance with the indications given in example 1. The filtrate is collected in accordance with example 2 and the DNA is extracted according to the extraction process according to the invention. 
         [0335]    The filtrate treatment protocol is identical to that mentioned in example 3 for genetic material. However, the eluate thus obtained is then sequenced using a new-generation PGM sequence (IonTorrent) with a 318 chip (reference: see table 7 below). 
         [0000]    
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 Reagents/kits used for sequencing the 
               
               
                 microbiota 
               
             
          
           
               
                 Reagents/kits 
                 Storage 
                 Supplier 
                 Ref. 
                 Batch 
               
               
                   
               
               
                 Ion PGM 
                 RT 
                 Life 
                 4482003 
                 057A03- 
               
               
                 Sequencing 
                   
                 Technologies 
                   
                 13 
               
               
                 Supplies 400 
               
               
                 Ion PGM 
                 −20° C. 
                 Life 
                 4482004 
                 057A03- 
               
               
                 Sequencing 
                   
                 Technologies 
                   
                 13 
               
               
                 Reagents 400 
               
               
                 Ion PGM 
                  +4° C. 
                 Life 
                 4482005 
                 057A03- 
               
               
                 Sequencing 
                   
                 Technologies 
                   
                 13 
               
               
                 Solutions 400 
               
               
                 Ion 318 Chip 
                 RT 
                 Life 
                 4484354 
                 P30756-1 
               
               
                 v2 kit 
                   
                 Technologies 
               
               
                   
               
             
          
         
       
     
         [0336]    In order to test the reproducibility of the protocol implementing the present invention, three repetitions per experimental condition were carried out so as to make it possible to calculate the sampling variability (CV), corresponding to all of the steps implemented by the protocol. 
         [0337]    The CV was calculated over the 10 phyla predominantly present in the stools. The results, presented in table 8 below, show that the CVs are equivalent between the two protocols (10-14%), with however a strong variation in value between phylum for the same protocol (6-23% for the process according to the invention and 2-35% for QiaGen). Consequently, this experiment shows that the DNA analysis process according to the invention makes it possible to obtain results that are equivalent in terms of reproducibility, while at the same time being simpler to produce than the process according to the QIAamp® DNA stool kit. 
         [0000]    
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 8 
               
             
             
               
                   
               
               
                 CV of the 10 phyla predominantly present in the stools 
               
             
          
           
               
                   
                   
                   
                   
                 Process 
               
               
                   
                   
                   
                   
                 according 
               
               
                   
                   
                   
                   
                 to the 
               
               
                   
                   
                   
                 QiaGen 
                 invention 
               
               
                 Taxon No. 
                 Rank 
                 Taxon 
                 CV (%) 
                 CV (%) 
               
               
                   
               
             
          
           
               
                 1239 
                 Phylum 
                 Firmicutes 
                 2% 
                 1% 
               
               
                 976 
                 Phylum 
                 Bacteroidetes 
                 15% 
                 6% 
               
               
                 201174 
                 Phylum 
                 Actinobacteria 
                 2% 
                 21% 
               
               
                 1224 
                 Phylum 
                 Proteobacteria 
                 12% 
                 13% 
               
               
                 508458 
                 Phylum 
                 Synergistes 
                 6% 
                 15% 
               
               
                 544448 
                 Phylum 
                 Tenericutes 
                 14% 
                 23% 
               
               
                 203691 
                 Phylum 
                 Spirochaetes 
                 2% 
                 17% 
               
               
                 1117 
                 Phylum 
                 Cyanobacteria 
                 35% 
                 5% 
               
               
                 1090 
                 Phylum 
                 Chlorobi 
                 17% 
                 23% 
               
               
                 3201, 3202 
                 Phylum 
                 Fusobacteria 
                 22% 
                 17% 
               
               
                 66 
               
               
                   
                   
                 Mean 
                 13% 
                 14% 
               
               
                   
                   
                 CV (CV 
                 82% 
                 55% 
               
               
                   
                   
                 variation) 
               
               
                   
               
             
          
         
       
     
       Example 7—Comparison in Terms of Quantification of Adenovirus DNA by PCR Using the Adenovirus R-Gene® Kit (Reference: 69-010B) after DNA Extraction with a) the QIAamp® DNA Stool Extraction Kit and with b) the Process for Extracting Biological Information According to the Invention 
       [0338]    In order to verify the concentration of adenovirus contained in the samples tested, stools containing a known concentration of adenovirus (“positive stool”) are diluted in a mixture of stools not containing adenovirus. The dilution of the positive stools is carried out in cascade to the following concentrations: 10 4 , 10 5 , 10 6  and 10 8  copies of virus per gram of stool. In the case of the stools containing adenoviruses, the Bristol types are generally between 4 and 7. 
         [0339]    Controls are systematically carried out:
       2 negative controls: non-inoculated stools detected as negative and non-inoculated phosphate EDTA buffer.       
 
         [0341]    For the adenovirus DNA extraction according to the QIAamp® DNA stool kit (ref. 51504, QiaGen), the supplier&#39;s experimental protocol is followed. 
         [0342]    The process for extracting biological information (adenovirus DNA) is described hereinafter. 
         [0343]    The 1) stool sampling, 2) suspending and 3) filtration steps are carried out in accordance with the teaching of example 2, using, as suspending solution, a solution consisting of phosphate (0.2 M), EDTA (50 mM), pH 8. 
         [0344]    The filtrate thus obtained is vortexed for a few seconds for the purposes of homogenization, and then 400 μl of homogenized filtrate are subsequently transferred into the wells of an easyMAG® shuttle (bioMérieux) and the “Dispense Lysis” program is launched in order to distribute 2 ml of lysis buffer (bioMérieux, reference: 280134). 
         [0345]    Once the “Dispense Lysis” program has ended, 10 μl of IC2 (internal control of the adenovirus R-Gene® kit), then 740 μl of a mixture, containing 600 μl of lysis buffer (bioMérieux, reference 280134) and 140 μl of silica, are added to each easyMAG® shuttle well. Once the mixture has been produced, the easyMAG® specific B, “off-board” lysis, program, elution in 50 μl, is launched. The eluate thus recovered is transferred into a new tube within 30 minutes following the end of the elution. 
         [0346]    A quantitative PCR analysis is carried out in order to quantify the adenovirus DNA extracted according to, on the one hand, the process according to the invention and, on the other hand, the process according to the QIAamp® DNA stool process, as a function of the theoretical amount initially inoculated into the suspending buffer. Thus, 10 μl of eluates are taken to be analyzed by quantitative PCR using the adenovirus R-Gene® kit. The results are expressed in log of the concentration of microorganisms inoculated. 
         [0347]    As shown by the results presented in  FIG. 26 , a linear relationship is noted between the number of viruses previously inoculated into the suspending buffer and the amount of DNA detected by quantitative PCR using the eluate obtained by implementing the process for extracting biological information according to the invention. Thus, the process for extracting biological information according to the invention (viral DNA) makes it possible to efficiently detect and quantify the adenoviruses initially present in a stool sample, and to obtain a result that is equivalent to the extraction process using the QIAamp® DNA stool kit, this being over a wide range of linearity (from 10 4  to 10 8  copies of virus per gram of stool). 
       Example 8—Test Showing the Amount of DNA Extracted from Samples of Stools of Different Bristol Type by Implementing the Process for Extracting Biological Information According to the Invention 
       [0348]    In order to evaluate the inter-stool reproducibility in terms of amount of DNA extracted from samples of stools of different Bristol types, several nucleic acid extractions were carried out on six different stool samples, the Bristol type of which is between 1 and 6, by implementing the process for extracting biological information (in the case in point DNA) according to the invention. 
         [0349]    The abovementioned process for extracting biological information according to the invention is identical to the process described in example 4 up to the step involving the easyMAG®. The eluates thus obtained are analyzed on a Nanodrop (ThermoScientific), in order to quantify and verify the purity (260/280 ratio and 260/230 ratio) of the DNA extracted. 
         [0350]    As illustrated in  FIG. 27 , a significant variation in the amount of DNA extracted is observed according to the Bristol type of the stool sample. Indeed, for stools of Bristol type 1 to 3, approximately 500 ng/μl of nucleic acids (5 μg of DNA) are extracted, while for stools of Bristol type 4 to 6, the nucleic acid concentration is close to 300 ng/μl nucleic acid (3 μg of DNA). The yield in terms of mass of DNA extracted is, consequently, approximately two times lower for the stools of Bristol type 4 to 6. However, the amount extracted remains sufficient to allow an analysis by PCR or by sequencing. 
         [0351]    This variation in terms of amount of DNA extracted is not due to the nature of the extraction process used, but is clearly due to the Bristol type of the stools. Indeed, a DNA extraction carried out using the Macherey Nagel kit (NucleoSpin Blood L), adapted for DNA extractions from liquid samples (for example from blood) is carried out on the stools of Bristol type 5 and 6, and gives similar results (results not represented). This phenomenon is explained by the fact that the stools of Bristol type 5 and 6 are more liquid—and thus by definition more dilute—which results in a decrease in the concentration of microorganisms and thus, de facto, in their DNAs. 
         [0352]    In conclusion, the process for extracting biological information (in the case in point DNA) according to the invention, using the device according to the invention, makes it possible to extract a sufficient amount of DNA from microorganisms in stools of Bristol type 1 to 6 to allow a subsequent analysis by PCR or sequencing, this being despite the variations observed in  FIG. 27 . 
         [0353]    It should be noted that the process for extracting biological information (in the case in point DNA) according to the invention, using the device according to the invention, also works for stools of Bristol type 7; the absence of results for this Bristol type being simply due to the non-availability of stools of Bristol type 7 at the time this example was carried out. 
       Example 9—Test Demonstrating the Efficiency of the Device According to the Invention for Taking and Analyzing Bovine Stool Samples 
       [0354]    The taking and analyzing of a bovine stool sample are carried out by means of the device of which the assembly is described in example 1, with the exception that:
       the calibrated sampling means  613  (comprising three calibrated hollow parts  6132 ,  6134  and  6135 ) of the device represented in  FIG. 16  is used, in order to collect 3000 mg of bovine stools, and   the volume of the TE buffer is increased to 10 ml.       
 
         [0357]    The device according to the invention was used as indicated in example 2 up to the filtration step inclusive. 
         [0358]    The device according to the invention thus made it possible to obtain 2 ml of a bovine stool filtrate, this being without clogging (blocking) of the filters of said device. 
         [0359]    In conclusion, with a few adjustments, the device according to the invention is entirely suitable for one or more veterinary application(s), for example in order to obtain all or part of the microorganisms present in a stool sample of animal origin, for example a bovine stool sample. 
       Example 10—Comparison of the Implementation Time and the Practicality of Use Between the Process for Extracting Biological Information According to the Invention and the QIAamp® DNA Stool Kit (QiaGen) 
       [0360]    In order to compare the implementation time and the practicality of use between the process for extracting biological information (in the case in point DNA) according to the invention and the protocol using the QIAamp® DNA stool kit, various DNA extractions were performed and the time for carrying out each step was reported in tables 9, 10 and 11 below, according to the type of process used. 
         [0361]    The process using the QIAamp® DNA stool kit was used to treat five stool samples. A total of 25 manual steps (including 15 sample preparation steps) and a total duration of 2h 20 (including 1h 57 for the sample preparation) were required. The various steps of the process using the QIAamp® DNA stool kit and their implementation times are presented in table 9 below: 
         [0000]    
       
         
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                   
                 TABLE 9 
               
             
             
               
                   
                   
               
               
                   
                 For 5 devices 
               
             
          
           
               
                   
                   
                   
                 Cumulative 
               
               
                   
                 Step of the “QIAamp ® 
                 Duration 
                 duration 
               
               
                 Item No. 
                 DNA stool” process 
                 (h:min) 
                 (h:min) 
               
               
                   
               
             
          
           
               
                 1 
                 Reagent and consumable 
                 00:09 
                   
               
               
                   
                 preparation 
               
               
                 2 
                 200-250 mg stool sample 
                 00:10 
                 00:10 
               
               
                   
                 taken 
               
               
                 3 
                 Lysozyme preparation 
                 00:10 
                 00:20 
               
               
                 4 
                 Lysozyme addition 
                 00:06 
                 00:26 
               
               
                 5 
                 Incubation 37° C. 
                 00:30 
                 00:56 
               
               
                 6 
                 Addition ASL + vortex 
                 00:05 
                 01:01 
               
               
                   
                 15 s/sample 
               
               
                 7 
                 Addition beads + 
                 00:08 
                 01:09 
               
               
                   
                 Vibrobeating 
               
               
                 8 
                 Incubation 95° C. + 
                 00:10 
                 01:19 
               
               
                   
                 preparation 2 ml tubes 
               
               
                 9 
                 Vortex 15s/sample. + 
                 00:06 
                 01:25 
               
               
                   
                 centrifugation + 
               
               
                   
                 transfer supernatant 
               
               
                 10 
                 Addition inhibitex + 
                 00:04 
                 01:29 
               
               
                   
                 vortex + incubation RT 
               
               
                 11 
                 Centrifugation 
                 00:06 
                 01:35 
               
               
                 12 
                 Transfer supernatant + 
                 00:05 
                 01:40 
               
               
                   
                 centrifugation 
               
               
                 13 
                 Preparation tubes + PK 
                 00:01 
                 01:41 
               
               
                 14 
                 Transfer supernatant + 
                 00:04 
                 01:45 
               
               
                   
                 addition AL + 
               
               
                   
                 homogenization 
               
               
                 15 
                 Incubation at 70° C. 
                 00:10 
                 01:55 
               
               
                 16 
                 Addition EtOH + vortex 
                 00:02 
                 01:57 
               
               
                 17 
                 Depositing on the 
                 00:02 
                 01:59 
               
               
                   
                 column 
               
               
                 18 
                 Centrifugation 
                 00:02 
                 02:01 
               
               
                 19 
                 Addition AW1 + 
                 00:02 
                 02:03 
               
               
                   
                 centrifugation 
               
               
                 20 
                 Addition AW2 + 
                 00:04 
                 02:07 
               
               
                   
                 centrifugation 
               
               
                 21 
                 Addition AW2 + 
                 00:05 
                 02:12 
               
               
                   
                 centrifugation 
               
               
                 22 
                 Addition AE 
                 00:01 
                 02:13 
               
               
                 23 
                 Incubation 
                 00:05 
                 02:18 
               
               
                 24 
                 Centrifugation 
                 00:01 
                 02:19 
               
               
                 25 
                 Preservation of eluate 
                 00:01 
                 02:20 
               
               
                   
               
             
          
         
       
     
         [0362]    Moreover, the process for extracting biological information (in the case in point viral DNA and bacterial DNA) according to the invention was also carried out in order to also treat five stool samples. 
         [0363]    The viral DNA extraction process according to the invention—for five samples—comprises eight steps (including three sample preparation steps) and is carried out in barely 1h 30 (including 30 minutes of sample preparation). The various steps of the viral DNA extraction process and their implementation times are reported in table 10 below: 
         [0000]    
       
         
               
               
             
               
               
               
               
             
           
               
                   
                 TABLE 10 
               
             
             
               
                   
                   
               
               
                   
                 For 5 devices 
               
             
          
           
               
                   
                   
                   
                 Cumulative 
               
               
                   
                 Step of the “viral DNA” 
                 Duration 
                 duration 
               
               
                 Item No. 
                 process 
                 (h:min) 
                 (h:min) 
               
               
                   
               
               
                 1 
                 Reagent and consumable 
                 00:14 
                 NA 
               
               
                   
                 preparation 
               
               
                 2 
                 Initiation of easyMAG ® 
                 00:13 
                 NA 
               
               
                 3 
                 200-250 mg stool sample 
                 00:04 
                 00:04 
               
               
                   
                 taken 
               
               
                 4 
                 Suspending 
                 00:05 
                 00:09 
               
               
                 5 
                 Filtration 
                 00:06 
                 00:15 
               
               
                 6 
                 Preparation easyMAG ® 
                 00:10 
                 00:25 
               
               
                   
                 shuttle 
               
               
                 7 
                 Launch easyMAG ® 
                 00:05 
                 00:30 
               
               
                   
                 protocol 
               
               
                 8 
                 Extraction in easyMAG ® 
                 01:00 
                 01:30 
               
               
                   
               
             
          
         
       
     
         [0364]    The bacterial DNA extraction process—for five samples comprises 12 steps (including seven sample preparation steps) and is carried out in 2h 13 (including 58 minutes of sample preparation). The various steps of the bacterial DNA analysis process and their implementation time are reported in table 11 below: 
         [0000]    
       
         
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                   
                 TABLE 11 
               
             
             
               
                   
                   
               
               
                   
                 For 5 devices 
               
             
          
           
               
                   
                   
                   
                 Cumulative 
               
               
                   
                 Step of the “bacterial 
                 Duration 
                 duration 
               
               
                 Item No. 
                 DNA” process 
                 (h:min) 
                 (h:min) 
               
               
                   
               
             
          
           
               
                 1 
                 Reagent and consumable 
                 00:14 
                 NA 
               
               
                   
                 preparation 
               
               
                 2 
                 Initiation of easyMAG ® 
                 00:13 
                 NA 
               
               
                 3 
                 200-250 mg stool sample 
                 00:04 
                 00:04 
               
               
                   
                 taken 
               
               
                 4 
                 Suspending 
                 00:05 
                 00:09 
               
               
                 5 
                 Filtration 
                 00:06 
                 00:15 
               
               
                 6 
                 Centrifugation 
                 00:06 
                 00:21 
               
               
                 7 
                 Pellet taken up and 
                 00:12 
                 00:33 
               
               
                   
                 resuspended 
               
               
                 8 
                 Mechanical lysis 
                 00:21 
                 00:54 
               
               
                 9 
                 Transfer supernatant 
                 00:04 
                 00:58 
               
               
                 10 
                 Preparation easyMAG ® 
                 00:10 
                 01:08 
               
               
                   
                 shuttle 
               
               
                 11 
                 Launch easyMAG ® 
                 00:05 
                 01:13 
               
               
                   
                 protocol 
               
               
                 12 
                 Extraction in easyMAG ® 
                 01:00 
                 02:13 
               
               
                   
               
             
          
         
       
     
         [0365]    In the light of the data presented in tables 9, 10 and 11 above, it appears that the bacterial DNA and viral DNA extraction processes according to the invention, using the device of the invention, make it possible to limit the number of steps and to significantly reduce the sample preparation time. 
         [0366]    Conversely, the process using the QIAamp® DNA stool kit requires a high number of manual and technically difficult steps, requiring the presence and continuous attention of a qualified laboratory technician. 
         [0367]    In conclusion, the device according to the invention makes it possible to simplify the experimental protocols (“viral DNA” and “bacterial DNA”) compared with the protocols of the prior art (QIAamp® DNA stool, QiaGen (ref. 51504)), by grouping together in one and the same device the elements required to carry out the sampling, suspending and filtration steps. This simplification makes it possible to significantly reduce the number and complexity of the manipulations, and to limit the risks of errors and of cross contaminations, while at the same time conferring increased working comfort for the normally qualified laboratory technician. 
       LITERATURE 
       [0000]    
       
         {1} Srijan A. Bodhidatta I., Mason C, Bunyarakyothin G, Jiarakul W, Vithayasai N. Field Evaluation of a Transport Medium and Enrichment Broth for Isolation of  Campylobacter  Species from Human Diarrheal Stool Samples. J Med Microbiol, 2013; 3, 48-52. 
         [2] Wasfy M, Oyofo B, Elgindy A. Churilla A. Comparison of preservation media for storage of stool samples. J Clin Microbiol 1995; 33:2176.