Sample withdrawal and dispensing device

A device is provided for receiving a fluid sample. The device includes a fitment having a cavity formed therein. The cavity is provided under vacuum. The fitment also includes a port having a seal. The port is configured to provide fluid connection from an exterior surface of the fitment to the cavity upon opening of the seal. The device optionally includes a collapsible compartment coupled to the fitment and in fluid communication with the cavity.

BACKGROUND AND SUMMARY OF THE INVENTION

The process of performing a chemical or biochemical analysis on a sample often involves a series of manual measuring and transfer motions, for example opening a container, dispensing the reagent solution, drawing a predefined amount of sample from the specimen, and so on. Serial manual volumetric measurements, multiple dispensing actions, and multiple opening and closing of containers are potential points of human error, contamination, and in some case, health risk. These potential problems are particularly acute for sample collection and analysis that need to occur outside of the controlled environment of the laboratory, such as, for example the collection of environmental samples for the detection of pathogens, infectious organisms, toxins, and bio-terrorism agents, as well as of forensic samples for the detection of human identifiers carried in the DNA, and the like.

In the field of clinical diagnostics, some of these concerns are addressed by the use of air evacuated tubes, such as VACUTAINER® tubes (Becton Dickinson and Company, of Rutherford, N.J.), for the collection of blood samples. These air evacuated tubes have needle penetrable stoppers inserted therein, and prevent the blood samples from becoming contaminated. The volume of blood to be withdrawn is controlled by the amount of vacuum in the tube, usually adjusted to partially fill the tube with blood. Evacuated glass vials prepackaged with reagents are described in U.S. Pat. No. 3,873,271, which also describes admitting a sample inside the evacuated vial by a cannula mounted in a special receptacle adapted to receive the vial. The use of one or more vacuum containers, or test tubes, to withdraw sample from a single syringe is described in U.S. Pat. No. 5,097,842. Further, in clinical diagnostics, robotics and automation are applied to withdraw blood samples from VACUTAINER® or other air evacuated containers, and dispense predetermined amounts of blood into reaction mixtures for analysis. These automation devices are often fairly large and may be unwieldy for sample collection and analysis outside of the laboratory.

According to one aspect of the present disclosure, a device for receiving a fluid is provided. The device includes a fitment and a collapsible compartment coupled to the fitment. The collapsible compartment is in fluid communication with the fitment. The fitment includes a cavity formed therein, and the cavity provided under vacuum. The fitment also includes a port having a seal. The port is configured to provide fluid communication from an exterior surface of the fitment to the cavity upon opening of the seal.

Illustratively according to this aspect of the disclosure, the cavity of the device is provided with a predetermined volume and a predetermined level of vacuum to receive a predetermined volume of the fluid upon opening of the seal. The device further includes a plunger sized to be received within the cavity. Activation of the plunger forces the predetermined volume of the fluid into the collapsible compartment.

Further illustratively, the plunger includes a notch configured to provide fluid communication between the cavity and the port when the notch is adjacent the port. The plunger acts to prevent fluid communication between the cavity and the port when the notch is rotated away from the port.

Additionally illustratively, the device further includes a dried reagent which may be contained within the collapsible compartment, the cavity, or both the collapsible compartment and the cavity. The dried reagent contained within the cavity may be the same as or different from the dried reagent contained within the compartment.

According to another aspect of the disclosure, a device for receiving a fluid is provided. The device includes a fitment having a plurality of cavities formed therein. Each cavity is provided under vacuum. The fitment further includes a channel fluidly connecting the cavities, a port extending from the channel to a surface of the fitment, and a seal provided at the port. The seal is configured to maintain vacuum in the cavities.

Illustratively according to this aspect of the disclosure, the seal may be breakable or the seal may be a unidirectional valve.

Further illustratively, the cavities are provided with a predetermined volume and a predetermined amount of vacuum such that upon opening the seal a predetermined volume of the fluid is drawn into each of the cavities. The device further includes means for sealing the fluid in each of the cavities.

Additionally illustratively, the device includes a plurality of collapsible compartments affixed to the fitment. Each collapsible compartment is in fluid communication with its respective cavity.

Further illustratively, the fitment further includes a plurality of additional cavities formed therein. Each additional cavity is provided under vacuum. The fitment further includes an additional channel fluidly connecting the additional cavities, an additional port extending from each additional channel to the surface of the fitment, and an additional seal provided at each additional port. The additional seal is configured to maintain vacuum in the additional cavities. The device further includes an additional plurality of collapsible compartments affixed to the fitment.

Each additional collapsible compartment is in fluid communication with its respective additional cavity.

Illustratively, the device further includes a plurality of plungers. Each plunger is sized to be received within its respective cavity. Activation of one of the plungers forces fluid received in the respective cavity into the collapsible compartment.

Further illustratively, a removable comb of the device may be provided to engage the plungers and normally prevent activation of the plungers.

Additionally illustratively, the channel of the fitment may be etched into the surface of the fitment and covered with a barrier material.

Further illustratively, the seal of the fitment includes a punctureable portion of the barrier material.

The plurality of cavities may form a row of cavities and the fitment may further include a plurality of additional rows of cavities. Each additional cavity is provided under vacuum. The fitment may further include a plurality of additional channels and a plurality of additional ports. Each additional channel connects the cavities of a respective row of cavities. Each additional port extends from a respective channel to the surface of the fitment. The fitment further includes a plurality of additional seals. Each seal is provided at its respective port and each additional seal is configured to maintain vacuum in its respective row of additional cavities. A removable cover may be provided to cover each cavity for maintaining vacuum within the cavities. Removal of the cover exposes the cavities to surrounding atmosphere.

According to yet another aspect of the present disclosure, a device for receiving a fluid sample includes a fitment and a flexible compartment coupled to the fitment. The fitment includes a vacuum chamber configured to maintain a vacuum therein and receive the fluid sample therein, a port in communication with the vacuum chamber and configured to receive the fluid sample therethrough, and a seal blocking the port. The flexible compartment is formed to define an interior region in fluid communication with the vacuum chamber. The flexible compartment is configured to receive the fluid sample therein.

According to this aspect of the present disclosure, the seal of the fitment is frangible. Further, the fitment is made of a generally non-compressible polymer material. The flexible compartment is made of a polymer.

According to another aspect of this disclosure, the device further includes a plunger received within the vacuum chamber and movable within the vacuum chamber to adjust a volume of open space unoccupied by the plunger within the vacuum chamber. The illustrative plunger includes a first end having a notch formed therein for alignment with the port of the fitment.

According to still another illustrative aspect of this disclosure, the fitment further includes a second port in communication with the vacuum chamber and configured to communicate with the surrounding atmosphere.

According to another aspect of the present disclosure, a device is configured to maintain an air-evacuated space therein and is provided for drawing a fluid sample into the air-evacuated space. The device includes a fitment and a flexible compartment coupled to the fitment. The fitment includes a vacuum chamber configured to maintain a vacuum therein, a first passageway in communication with the vacuum chamber and configured to communicate with the surrounding atmosphere, a second passageway in communication with the vacuum chamber and configured to communicate with the surrounding atmosphere, and a frangible seal positioned to block the second passageway to prevent communication between the vacuum chamber and the surrounding atmosphere. The flexible compartment of the device is formed to define an interior region configured to receive the fluid sample therein. The interior region is positioned in fluid communication with the vacuum chamber. The device further includes a plunger received within the vacuum chamber for up and down movement within the vacuum chamber to adjust a volume of open space unoccupied by the plunger within the vacuum chamber.

According this aspect of the disclosure, the plunger includes a notch for alignment with the second passageway of the fitment. The illustrative plunger is movable between a first position to block communication between the vacuum chamber and the first passageway and a second position to block communication between the vacuum chamber and the second port.

Further illustratively according to this aspect of the disclosure, the first passageway is less than 1 mm in diameter, the second passageway is less than 1 mm in diameter, and the vacuum chamber is 5 mm in diameter.

Additionally illustratively according to this aspect of the disclosure, the flexible compartment is made of a polyvinyl material. The fitment is made of a soft polymer plastic material and the plunger is made of a rigid polymer plastic material. Further, a diameter of the plunger is substantially equal to a diameter of the vacuum chamber.

Further illustratively according to this aspect of the disclosure, the air-evacuated space has a predetermined volume and is provided with a predetermined level of vacuum for drawing in a predetermined volume of the fluid sample.

According to yet another aspect of the disclosure, a pouch assembly for receiving multiple fluid samples therein is provided. The pouch assembly includes a fitment and a plurality of flexible compartments coupled to the fitment. The fitment includes a plurality of vacuum chambers formed therein, a sample access port in communication with at least one of the plurality of vacuum chambers, and a plurality of vacuum holes. Each vacuum hole is in fluid communication with one of the plurality of vacuum chambers. Each flexible compartment of the plurality of compartments is in fluid communication with one of the plurality of vacuum chambers.

Illustratively according to this aspect of the disclosure, the sample access port is in communication with each of the plurality of vacuum chambers. The fitment further includes a passageway between the sample entry port and each of the plurality of vacuum chambers. Further illustratively, the sample access port is a plurality of sample access ports and further each sample access port is in fluid communication with one of the plurality of vacuum chambers.

Still according to another aspect of the disclosure, a method of introducing a pre-measured amount of a fluid sample into a pouch assembly is provided. The pouch assembly includes a flexible compartment and a fitment coupled to the flexible compartment. The fitment includes a vacuum-evacuated cavity in fluid communication with the flexible compartment. The method includes breaking a seal of the fitment to provide communication between the vacuum evacuated cavity and the fluid sample, allowing the fluid sample to be drawn into the cavity, and moving the fluid sample from the cavity into the flexible compartment.

Illustratively according to this aspect of the present disclosure, moving the fluid sample from the cavity into the flexible compartment includes moving a plunger positioned within the cavity to push the fluid sample from the cavity into the flexible compartment.

Further illustratively according to this aspect of the present disclosure, the method further includes the step of creating a vacuum in the cavity by placing the pouch assembly in a vacuum chamber and evacuating air from within pouch assembly through a vacuum port of the fitment. The vacuum port is in communication with the cavity. Further, the step of creating the vacuum occurs prior to the step of breaking the seal. Still further, the step of creating the vacuum further includes plugging the vacuum port once the vacuum within the cavity is approximately 7 Pa. The step of creating the vacuum further includes plugging the vacuum port by moving a plunger of the pouch assembly within the cavity to block communication between the vacuum port and the cavity. The step of creating the vacuum may further illustratively include moving a plunger of the pouch assembly within the cavity to adjust a volume of open space of the cavity unoccupied by the plunger.

According to still another aspect of the present disclosure, a method of manufacturing a pouch assembly including a flexible compartment and a fitment coupled to the flexible compartment for receiving a predetermined amount of fluid sample therein is provided. The method includes molding the fitment of the pouch assembly from a polymer plastics material to include a vacuum cavity, etching a plurality of channels into a first surface of the fitment for communication with the vacuum cavity of the fitment, and coupling a flexible compartment of the pouch assembly to the fitment. The flexible compartment is in fluid communication with the vacuum cavity.

Illustratively according to this aspect of the disclosure, the coupling step includes coupling a top layer of the flexible compartment to the first surface of the fitment to cover the plurality of channels etched into the first surface and coupling a bottom layer of the flexible compartment to a second surface of the fitment to cover an aperture of the cavity formed therein. Further illustratively, coupling the top layer of the flexible compartment includes heat sealing the top layer to the first surface; coupling the bottom layer of the flexible compartment includes heat sealing the bottom layer to the second surface of the fitment.

Additional features of the present invention will become apparent to those skilled in the art upon consideration of the following detailed description of preferred embodiments exemplifying the best mode of carrying out the invention as presently perceived.

DETAILED DESCRIPTION

A pouch assembly10, shown inFIGS. 1A and 1B, is provided for receiving a fluid sample56(shown inFIGS. 2C-2E). Illustrative pouch assembly10includes a collapsible compartment12, a fitment14coupled to the collapsible compartment12, and a plunger16received within a cavity18of the fitment14. As shown inFIG. 1A, collapsible compartment12includes a top layer20coupled to a front surface22of the fitment14and a bottom layer24coupled to a portion of a bottom surface26of the fitment14. Illustratively, top and bottom layers20,24are formed of a barrier material (defined below) which has been folded in half to create a bottom or end28of compartment12as well as top and bottom layers20,24of compartment12. Top and bottom layers20,24are coupled to each other, as is discussed in greater detail below, to form compartment12having an interior region30for receiving fluid samples therein, for example.

The fitment14, as mentioned above, includes cavity18. Cavity18is in communication with interior region30of compartment12as shown inFIG. 1A. Illustrative cavity18of fitment14is cylindrical in shape and is formed to extend from a top surface32of fitment14to bottom surface26. Illustrative cavity18is defined by an interior surface34of fitment14and has a diameter of approximately 5 mm; however, it is within the scope of this disclosure to include a cavity having other suitable diameters. For example, the cavity diameter may vary depending upon the volume of sample fluid desired to be deposited within each compartment. A diameter of a corresponding plunger16for use within cavity18may approximately the same or slightly larger than the cavity diameter in order to maintain a tight seal to provide a press-fit or interference-fit with the cavity18. Illustratively plunger16is sized to slide within cavity18with a force of between approximately 1 to 20 N.

A vacuum port36of fitment14is formed through a rear surface38of fitment14to communicate with cavity18along a channel40. Illustrative port36is approximately 2 mm in diameter; however, it is within the scope of this disclosure to include a vacuum port having other suitable diameters. As is discussed in more detail below, vacuum port36is provided for communication with a vacuum or vacuum chamber (not shown) to draw out the air from within pouch assembly10to create a vacuum within cavity18and interior region30of compartment12.

Illustrative fitment14further includes a sample entry port42formed in the rear surface38of fitment14. Illustratively, sample entry port42is positioned below vacuum port36, as shown inFIGS. 1A and 1B. A channel44of fitment14is formed between cavity18and sample entry port42. A seal46of fitment14is positioned within channel44to normally prevent communication between cavity18and the surrounding atmosphere via channel44and sample entry port42.

As is discussed in greater detail below, seal46is frangible and may be broken upon insertion of a cannula (not shown), for example, through sample entry port42in order to allow a fluid sample from within the cannula to be drawn into cavity18. Illustrative seal46is made of the same material as fitment14. However, it is within the scope of this disclosure for the seal to be made of other suitable materials, such as, rubber, thin plastic film, and other elastomers, for example. Further, it is within scope of this disclosure for the seal to be positioned anywhere along channel44or within cavity18, or covering port42to block communication between the cavity18and the surrounding atmosphere. In other words, port42is essentially a sealed port. Illustratively, sample entry port42is approximately equal to or less than 1 mm in diameter and channel44is similarly approximately equal to or less than 1 mm in diameter. However, it is within the scope of this disclosure to include a sample entry port and connecting channel having other suitable dimensions.

The illustrative plunger16of the pouch assembly10is cylindrical in shape and has a diameter of approximately 5 mm to be press-fit into cavity18. Plunger16includes a first end portion48and an opposite second end portion50. A notch52of plunger16is formed in second end portion50, as shown in FIGS.1A and2A-2E, for example. In use, second end portion50is inserted into cavity18at top surface32of fitment14. As is discussed in more detail below, notch52may be aligned with sample entry port42to allow a fluid sample to be drawn into cavity18, as shown inFIG. 2C.

In describing the invention, the following terminology will be used in accordance with the definitions set forth below.

As used herein, the term “barrier material” refers to the flexible material from which the collapsible compartment12of the pouch assembly10is illustratively constructed. The barrier material potentially may be a single layer or a laminated structure, and, depending on the application, is preferably air and water impermeable. Other characteristics of the barrier material are dictated by the conditions of storage prior to use, the conditions during use, the nature of material that is to be contained in the collapsible compartment12, and the nature of reaction and interrogation that is to be performed on the contained material. For instance, if the reaction is to be monitored optically, then at least a portion of the barrier material should be optically clear to the excitation and emission wavelengths used. If PCR is to be used, the barrier material should be able to withstand temperature cycling. Exemplary barrier materials include, but are not limited to, polyester, polyethylene terephthalate (PET), polycarbonate, polypropylene, polymethylmethacrylate, and mixtures thereof, and can be made by any process known in the art, including extrusion, plasma deposition, and lamination. Metal foils or plastics with aluminum lamination also may be used. Other barrier materials are known in the art. In an illustrated embodiment, for use with PCR, the collapsible compartment has a coefficient of heat transfer of approximately 0.02 to 20 W/m*degK.

If fluorescence monitoring of a reaction is desired, plastic films that are adequately low in absorbance and auto-fluorescence at the operative wavelengths are preferred. Such material could be identified by trying different plastics, different plasticizers, and composite ratios, as well as different thicknesses of the film. For plastics with aluminum or other foil lamination, the portion of the collapsible compartment12that is to be read by a fluorescence detection device can be left without the foil. For example, if fluorescence is monitored through the bottom28of pouch assembly10, then bottom28would be left without the foil. In the example of PCR, film laminates composed of polyester (Mylar, Dupont, Wilmington Del.) of about 0.0048 inch (0.1219 mm) thick and polypropylene films of 0.001-0.003 inch (0.025-0.076 mm) thick perform well. Illustratively, each layer20,24of collapsible compartment12is made of a clear material so that the collapsible compartment12is capable of transmitting approximately 80%-90% of incident light.

The term “flexible” is herein used to describe a physical characteristic of the barrier material of the collapsible compartment12. The term “flexible” is herein defined as readily deformable and collapsible by the levels of vacuum used without cracking, breaking, crazing or the like, and readily returned essentially to the non-collapsed state with ease. For example, thin plastic sheets, such as Saran wrap and Ziplock bags, as well as thin metal foil, such as aluminum foil are flexible. Standard thin glass capillaries with outer diameter of about 1 mm may flex with attempts to bend, however, they are not flexible within the above-referenced definition.

The term “vacuum” refers to a pressure below atmospheric pressure. In illustrative examples of vacuum of 240 Pa or less or 7 Pa or less is used. However, other levels of reduced pressure are within the scope of this disclosure.

When reference is made to sealing the barrier material to itself, or to the material used for the non-collapsible fitment14, the method may be chosen from one of many known in the art. Illustratively, the seal is tight enough to endure force of vacuum down to 100 Pa or pressure up to 40 psi, more preferably, down to 50 Pa or even 20 psi, and most preferably down to 5 Pa. Heat sealing is one of the more commonly used methods, whereby heat is used to fuse the barrier material to itself or to different materials and thereby form a seal. For example, as shown inFIG. 1B, seams54are created by heat sealing top layer20to bottom layer24. Interior region30is defined between spaced-apart seams54and bottom28of collapsible compartment12. Adhesive joining may also be used, whereby an adhesive is applied to one or both layers20,24to be sealed prior to sealing. Welding techniques, such as radio frequency welding and ultrasonic welding also may be used depending on the barrier material and other materials to which the barrier material needs to adhere. Infrared can in some cases be used to seal barrier material. Other methods of sealing a pouch are known in the art and are within the scope of this disclosure.

When reference is made to fitment14, the term is used to describe a non-collapsible part of the pouch assembly10. The term non-collapsible is herein used in reference to the fitment14and the ability of fitment14to withstand certain negative pressures applied thereon without substantially collapsing or deforming cavity18and/or other passageways formed within fitment14. Fitment14is constructed from material chosen from a variety of plastics, including the use of two or more different plastics to provide different characteristics for different parts of the fitment14. For example, the body of fitment14may be made of a rigid plastic having an elastomeric overmold. Illustratively, the fitment14should be firm enough so that cavity18will not significantly change volume under vacuum, but also soft enough so that seal46can be easily punctured by a cannula, or the like, leaving a relatively clean break in the puncture region so as not to release debris that can block the channel44through which fluids are introduced into the cavity18. For example, fitment14may be made of a material which collapses approximately 5-10% when under vacuum. However, in some embodiments a more flexible material may be used for the fitment, particularly if the fitment material flexes in a uniform manner and provides a uniform volume in the cavity. Although illustrative seal46is made from the same material as fitment14, it is within the scope of this disclosure to include a seal made of other materials such as, for example, the barrier material. Optionally, the material used in seal46is capable of self-sealing to minimize leakage or backflow. Furthermore, the material of the fitment14should adhere tightly to the barrier material by means of sealing, as described above.

Other characteristics of the fitment material are dictated by the storage and use conditions of the pouch assembly10, which can be selected by those skilled in the art. Illustratively, the fitment14may be manufactured from polypropylene. As mentioned above, however, fitment14may also be made of an elastomeric material or may be made of a more rigid plastic and an elastomeric overmold. Other suitable materials may be used. Further illustratively, the fitment14may be injection molded from a plastic material and the cavities and passageways of fitment14may be formed therein during the injection molding process. Alternatively, the cavities and passageways may be formed by machining after the injection molding process.

When reference is made to the plunger16, the term is used to describe a movable part that is inserted into cavity18of fitment14. The plunger16illustratively can be constructed from materials selected from a group of hard plastics, soft rubber, or soft plastics that will seal the fitment cavity18to hold vacuum. The choice of material of plunger16may depend on the fitment material, particularly the material used at a seal surface34of fitment14defining cavity18that will be in contact with the plunger16. If the seal surface34is a hard plastic, for example, then a soft rubber or soft plastic may be used as plunger material. Alternatively, if the seal surface34is a soft plastic, for example, then a hard plastic plunger material often will be appropriate, with use of vacuum grease (not shown), if desired, on the seal surface34. Furthermore, the plunger material should accommodate designs to prevent backflow when the plunger16is used to push fluid into the interior region30of one or more collapsible compartments12. For example, a plunger formed by injection molding, for example, may include a parting line formed where two injection mold components, for example, come together. This parting line of the plunger may lie along the seal surface34of cavity18and may permit the fluid sample to flow back up along the parting line of the plunger rather than into the compartments. In other words, a plunger having a smooth, uniform outer surface for engaging seal surface34of cavity18to form a seal may prevent backflow of the fluid sample as the fluid sample is moved to the compartment. Similarly, notch52of plunger allows incoming fluid through channel44to enter cavity18when notch52is aligned with channel44. The plunger16can then be rotated to block channel44when plunger16is fully depressed to move the fluid within cavity18to the compartment12without allowing the fluid to move back into channel44.

As mentioned above, in the embodiment shown inFIGS. 1A-2E, a barrier material is folded, and/or sealed to itself, on three sides generating the collapsible compartment12, leaving the fourth side open to be sealed to the fitment14. When the barrier material is sealed to a fitment3, collapsible compartment12is fluidly connected to the fitment cavity18. As shown, plunger16is partially inserted to a first position within the cavity18so as to leave open port36, and channel44, as shown inFIGS. 1A and 2A. Air is evacuated from the pouch assembly10, illustratively by placing the whole pouch assembly10into a vacuum chamber (not shown). Vacuum is applied and air within the pouch assembly10is evacuated through port36. The compartment12is collapsed at this point, with top and bottom layers20and24in full contact with each other, as shown inFIG. 2B.

Preferably a vacuum of 240 Pa or less is used, and most preferably a vacuum of 7 Pa or less is used. The length of time required for evacuating air depends on several factors including, but not limited to, the size of the pouch assembly, duration required to degas the barrier material and materials used for the fitment and plungers, air penetration and out-gassing rates of said materials, and the required shelf life of the pouch assembly. Typically, a pouch assembly, such as the pouch assembly shown inFIG. 1A, may be degassed under a moderate vacuum anywhere between 12 and 72 hours. The conditions of vacuum can be optimized by those skilled in the art.

After an appropriate amount of vacuum is applied, plunger16is lowered to a second position within cavity18where port36is blocked and a volume of space within cavity18unoccupied by plunger16is reduced to a predetermined volume. The final level of vacuum and volume of cavity18define a predetermined volume of fluid sample that will be drawn into cavity18, as shown inFIG. 2B. Said volume may be equal to or smaller than the fully inflated volume of compartment12, and may also depend on the actual biochemical or chemical reaction process to be performed. At this stage, channel44has access to cavity18; i.e. is not blocked by the plunger16. Illustratively, notch52formed in the plunger16provides this access, as shown inFIG. 2B. However, it is within the scope of this disclosure to include other means of providing access between port42and cavity18when plunger16is in the second position. For example, port42may be positioned further away from port36. An optional holding device can be used to secure the position of the plunger16, as shown, for example, inFIGS. 5-7Bas comb or separator470of a pouch assembly410discussed in greater detail below. For storage, the pouch assembly10optionally can be placed inside another vacuum evacuated pouch capable of holding a vacuum illustratively around 500 Pa. In an alternative embodiment, the pouch assembly optionally can be placed inside an air-evacuated air-tight non-collapsible container or alternatively, inside a pouch with an internal rigid frame or container that provides a non-collapsible space of vacuum large enough to hold the pouch assembly, and to maintain the vacuum inside the pouch assembly and keep compartments fully collapsed during long-term storage.

In using the air-evacuated pouch assembly10, illustratively, a fluid sample56is placed in a container (not shown) with a syringe having a cannulated tip that can be inserted into sample entry port42to puncture seal46therein. Alternatively, the fluid sample56may be withdrawn directly from its source through a cannula, or the like. When seal46is punctured, the fluid56is withdrawn from the container (or its source) due to the negative pressure within cavity18. Fluid56then passes through port42and channel44to fill cavity18, as shown inFIG. 2C. At this point, the fluid56usually does not enter collapsed compartment12. The fluid sample56can be liquid, gel, or gas as long as the fluid sample56is capable of being drawn into cavity18by force of vacuum. Finally, the plunger16is lowered to a third position within cavity18to lie at a bottom of cavity18generally flush with bottom surface26of fitment14, as shown inFIG. 2E, to push the fluid56into the flexible compartment12, where biochemical, or chemical reactions can take place, and analysis may be performed by optical or other means of interrogation.

If a plunger design is used including notch52, as illustrated in the embodiment shown inFIGS. 1A-2E, the plunger16may be rotated, as shown inFIG. 2D, prior to being lowered so as to offset notch52and to close off channel44from communication with cavity18. This acts to minimize any potential backflow of fluid through channel44to the surrounding atmosphere. As mentioned above, although notch52is shown and described above with respect to plunger16, it is within the scope of this disclosure to close off either channel44or sample entry port42soon after dispensing the fluid sample56into the cavity18by other means, such as depressing plunger16toward the bottom of cavity18, heat sealing, unidirectional valves, or self-sealing ports, for example.

Prior to use, reagents (not shown) may also be placed either in the cavity18or in the collapsible compartment12, or in both. The reagents may then be dried through the vacuuming process. A freeze-dryer or a lyophilizer can be used to apply vacuum. It is contemplated that after a fluid sample56is dispensed into the fitment14of such a pouch assembly10having reagents therein prior to injection of the fluid sample56, the fluid sample56is mixed with dried reagents in the fitment cavity18, and the resulting mixture is transferred to the collapsible compartment12. It is further contemplated that a first reaction may take place within the fitment cavity18, and a second reaction may take place within the flexible compartment12, particularly if cavity18and compartment12each contain different reagents.

In a further embodiment, shown inFIG. 3A, another illustrative pouch assembly110is provided. Pouch assembly110is similar to pouch assembly10, and therefore, like reference numerals have been used to identify like components. Pouch assembly110includes a row111of collapsible compartments12divided by seams54created by sealing the barrier material, as described above. Pouch assembly110also includes a fitment114coupled to row111of compartments12. Fitment114includes three cavities18spaced-apart from one another. Illustratively, row111includes three compartments12such that each compartment12is in communication with a corresponding cavity18of fitment114. Illustratively, each compartment-cavity combination is sealed from fluid communication with any other compartment or cavity. Therefore, the cavities18are in one-to-one communication with their respective compartments12.

Fitment114of pouch assembly110also includes three separate vacuum ports36spaced-apart from each other. Each vacuum port36is in communication with one of the three cavities18of fitment114. Fitment114further includes sample entry port42. An interior branched channel144of fitment114(shown in phantom inFIG. 3A) provides communication between sample entry port42and each of the three cavities18of fitment114. For example, channel144includes branches146which extend from a main passageway148of channel144to each cavity18. Seal46is provided within channel144near sample entry port42prior to communication with main passageway148of channel144. Pouch assembly110further includes three plungers16(not shown inFIG. 3A). Each plunger16is received within a corresponding cavity18of fitment114.

Multi-compartment pouch assembly110is air-evacuated in the same manner as that described above with respect to pouch assembly10. Once air has been evacuated and the respective plungers16have been depressed to their second position within cavity18to block each respective port36, a fluid sample (not shown) may be introduced. In using multi-compartment pouch assembly110, a sample is dispensed through the single sample entry port42to the multiple cavities18through branched channel144that is in communication with, and capable of distributing the sample to, said multiple cavities18, as shown inFIG. 3A. A substantially equal predetermined amount of the fluid sample is drawn into each respective cavity18. In this configuration, it may be important to optimize the dimension and design of the branched channel144to minimize diffusion and mixing of fluid between multiple cavities prior to closing off the access to channel144by plungers16or other means. In the example of a liquid sample, such design optimization of channel144can be achieved by use of different color dyes placed in each fitment cavity18, injecting a liquid similar or equal in density and viscosity to the sample, and following the diffusion and mixing rate of the colored liquid across cavities. In an exemplary operational design, diffusion and mixing of water-based liquid between multiple cavities are minimized when channel144has a square cross-section of about 1 mm along its entire path, when the path is approximately 1-3 cm per branched channel, and when the sealing event can be performed between 10 seconds to 10 minutes after injection of the liquid. Diffusion may further be minimized by either further decreasing the dimension of the channel, further increasing the distance between fitment cavities, or further decreasing the time between sample injection and sealing. Optionally, mixing may be prevented by using unidirectional valves at or near the junction between channel144and each fitment cavity18.

Yet another pouch assembly210is shown inFIG. 3B. Pouch assembly210is a multi-compartment pouch assembly similar to assembly110shown inFIG. 3A. Similarly, like reference numerals are used to identify like components. Pouch assembly210includes row111of compartments12coupled to a fitment214. Different from fitment114, fitment214includes three separate sample entry ports42formed therein as well as three separate channels44. Each channel44is in communication with one of the sample entry ports42and a corresponding cavity18. Similar to fitment14of pouch assembly10, a seal46is provided within each channel44. In using this multi-compartment pouch assembly210, each injected sample (not shown) is individually dispensed into single cavities18. Although not shown, three plungers16are provided in pouch assembly210. Each plunger16is to be received within one of the three cavities18as is described above with respect to the aforementioned embodiments.

In yet another embodiment, shown inFIGS. 4A and 4B, an alternative pouch assembly310is provided. Pouch assembly310includes a fitment314comprised of multiple cavities or wells318, illustratively, seven cavities318, connected by a channel344, similar to channel144shown inFIG. 3Awith respect to fitment114. Illustrative channel344includes single main passageway348formed to extend along a length320of fitment314and multiple branches346each connecting a cavity318with the main passageway348. Illustratively, therefore, there are seven branches346. Illustrative fitment314has a height322smaller than an illustrative height of the fitments14,114,214described above. A single sample entry port42of fitment314is formed in a top surface332of fitment314. Sample entry port42is in communication with main branch348of channel344. The seal46is provided between sample entry port42and main branch348.

In this embodiment, the pouch assembly310lacks the collapsible compartments and plungers described above with respect to pouch assemblies10,110, and210. Further, each cavity318is defined by a closed bottom surface315and an open top aperture316formed in a top surface332of fitment314. Reagents are placed into each of the cavities318through the open aperture316, and then dried or immobilized onto the interior surface317of the cavities318by methods known in the art. After evacuation of air, the open top aperture316is sealed with a material347(shown inFIG. 4C) so that an unoccupied space of each cavity318is now reduced to a predetermined volume. The material used to seal the open top316may be, for example, the same material as assembly310. Alternatively, a flexible barrier material may be used and may be attached to top surface332of fitment314by heat sealing, for example. The material used to seal the open top aperture316may also seal sample entry port42, to provide seal46.

When the seal46is punctured, and a fluid sample (not shown) is taken in, the fluid sample is distributed into each cavity318through channel344. After the fluid sample is dispensed into each cavity318, access from each cavity318to channel344may be closed by heat sealing or other means. Branches146of channel144may be heat sealed along line350, for example. If reagents are dried in the cavities318, then the dimensional design of channel344may be optimized to minimize diffusion of sample across cavities before said sealing event. Such design may include the use of narrower channels closer to the position of the seal46, as discussed above. Channel344can be embedded inside fitment314, or alternatively etched on the top surface332of fitment314. This etched channel (not shown) may be later covered when the cavities318are sealed at the open aperture316by barrier material or the like.

In a further embodiment, shown inFIG. 4C, a pouch assembly350is provided including a fitment352having a two-dimensional row of cavities318. Each row is provided with channel344, although other arrangements are contemplated. Each channel344is in communication with one sample entry port42formed in top surface332of fitment352. Although the illustrated embodiment shows an array of three rows of seven cavities, other arrangements are within the scope of this disclosure. For example, a pouch assembly350may be arranged like a microtiter plate, for example, a96,384, or1536well plate. Samples may then be processed using standard devices configured to receive microtiter plates. Optionally, cover347may be removable for further processing of the contents of cavities318.

As shown inFIGS. 5 and 6, fitment414includes a top surface432, a bottom surface426, a front surface422, a rear surface438, and end surfaces442and444. Fitment414further includes twelve cavities18spaced-apart from each other and each formed through fitment414to extend from top surface432to bottom surface426. Although illustrative fitment414includes twelve cavities18, it is within the scope of this disclosure to include a fitment having any suitable number of cavities formed therein.

In addition to fitment414, pouch assembly410further includes twelve plungers416each received within one of the corresponding cavities18, as shown inFIGS. 5 and 6. Each plunger416includes a top head portion450, an end portion452, and a central neck portion454positioned between and coupled to both the top head portion450and the end portion452. Illustrative head portion450is pentagonal in shape and includes a notch456formed in a top surface458of head portion450. Notch456is provided for use during optional automated filling of pouch assembly410.

Neck portion454includes a first end460coupled to the head portion450and a second end462coupled to the end portion452. Illustrative neck portion454has a smaller cross-sectional region or diameter than head portion450. Illustrative neck portion454is approximately 20 mm long. End portion452is coupled to second end462of neck portion454and is generally cylindrical in shape. A cross-sectional region or diameter of end portion452is slightly larger than the cross-sectional region of neck portion454. Thus, as illustrated, neck portion454is narrower than both head portion450and end portion452. Similarly, a diameter of end portion452is approximately 5 mm and the diameter of each cavity18is approximately 5 mm to ensure a press-fit between end portion452of plunger416and the sealing wall or interior surface34defining each cavity18. As discussed above with reference to plunger16, end portion452of plunger416similarly includes notch52formed therein, as shown inFIG. 7A.

In addition to the twelve compartment row111, fitment414, and plungers416, pouch assembly410further includes a comb or separator470normally positioned between top surface432of fitment414and head portion450of plunger416, as shown, for example, inFIGS. 5 and 6. Separator470acts as a lock to maintain the plungers416in a particular position relative to fitment414. Illustrative separator470is shaped like a comb and includes a connecting backbone472and multiple teeth474extending therefrom. Teeth474are spaced-apart from each other to define notches476of separator470each formed to receive a portion of the neck portion454of a respective plunger416therein. Illustrative comb470is divided into four comb portions, as shown inFIG. 5such that each comb portion communicates with three respective plungers. As is discussed in more detail below, this allows a user to operate only one set of three plungers at any desired time.

Looking now toFIG. 5, fitment414includes a vacuum port36associated with each cavity18. Thus, fitment414includes twelve vacuum ports36. Although fitment414includes a separate vacuum port36for each cavity18, it is within the scope of this disclosure to include a fitment having only one vacuum port36, for example, which is interconnected through a network of channels to each cavity18of fitment414. Fitment414further includes four sample entry ports478,480,482,484each formed through rear surface438of fitment414. First and second sample entry ports478,480are formed at a left end of fitment414(as shown inFIG. 5) while third and fourth sample entry ports482,484are formed at a right end of fitment414.

Fitment414further includes multiple channels in communication with sets of cavities18of fitment414, as shown inFIGS. 6,7A, and7B. A channel486communicates with third sample entry port482via a passageway488of channel486. Channel486further includes a main passageway490and three branches492. Each branch492provides communication between the main passageway490and a corresponding cavity18. The channel486thus operates similarly to channel144disclosed above with respect to pouch assembly110. Another channel494communicates with fourth sample entry port484via a passageway496of channel494. Channel494further includes a main passageway498and three branches500extending therefrom. Each branch500provides communication between the main passageway498and a corresponding cavity18.

Similarly, another channel502is provided to communicate with the first sample entry port478and includes a passageway504in direct communication with the port478as well as a main passageway506etched in front surface422of fitment414and three branches508connecting the main passageway506to three cavities18. Another channel510, similar to channel494, is provided to communicate with second sample entry port480and includes a passageway512in direct communication with the port480as well as a main passageway514etched in front surface422and three branches516connecting the main passageway514to three cavities18. Thus, via the system of channels486,494502,510each of the first, second, third, and fourth sample entry ports482,484,478,480is in fluid communication with three corresponding cavities18. Although main passageway490,498506,514of the channels486,494,502,510are etched in front surface422of fitment414, it is within the scope of this disclosure to provide channels or passageways formed through fitment414similar to the channels144of fitment114, for example. The illustrative main passageways490,498,506,514which are etched into front surface422of fitment416are sealed by a portion of bottom layer24of row111of compartments12, as shown inFIG. 6.

As mentioned above, fitment414includes four sample entry ports478,480,482, and484each with seals42positioned therein, which if broken will connect each port478,480,482,484with three corresponding cavities18via the branched channels486,494,502,510described above. As shown inFIG. 7A, plungers416are in a first position inserted partially in said cavities18to expose the vacuum port36to allow air to be evacuated from within pouch assembly410. Illustratively, after air is evacuated from the pouch assembly410, the volume of open space unoccupied by a respective plunger416within each cavity18is adjusted by lowering each plunger416to the second position shown inFIG. 5to block vacuum port36while leaving open access of each cavity to the respective sample entry ports478,480,482,484. The plungers416are locked or secured in this second position illustratively by comb470. When the seal42of one of the sample entry ports478,480,482,484is broken, the fluid sample is withdrawn by force of the vacuum from the three corresponding cavities18in communication with that particular sample entry port. In one embodiment, the diameter of channels486,494,502,510is kept small, such as, for example, approximately equal to or less than 1 mm, to minimize diffusion of fluid across cavities18.

Subsequently, the comb470or a portion of the comb470is disengaged from fitment414, and the respective unlocked plungers416are twisted to seal access (in the form of notch52) between each cavity18and respective channels486,494. The unlocked plungers416are then lowered to the bottom of cavity18to the third position to dispense the fluid sample into the three respective compartments12in communication with the three cavities18. Optionally, as mentioned above, comb470may be broken into multiple sections to secure a certain number of plungers416in the second position within each cavity18. Illustratively, comb470is broken into four sections. Each comb section includes three detents476for receiving a portion of three plungers416therein to lock or secure three plungers416in the second position. Thus, a three-plunger section of the comb470may be removed to activate three plungers416at one time while reserving the remaining plungers416for later use. Alternatively, comb470may be provided in a unitary piece for activation of all 12 plungers.

It is contemplated that the devices of the present disclosure may be used for testing multiple pathogens or multiple genes from a single source. As illustrated, the device ofFIGS. 4A and 4Bis configured for testing a single sample for seven items, while the device ofFIGS. 5-7Bis configured for performing three tests each on four samples. Other configurations are within the scope of this disclosure.

The following examples are given to illustrate various embodiments which have been made with the present invention. It is to be understood that the following examples are not comprehensive or exhaustive of the many types of embodiments which can be prepared in accordance with the present invention.

A twelve-compartment pouch assembly410(FIGS. 5-7B) is constructed from polyethylene terephthalate-polypropylene laminates (0.48 mill PET/2 mill polypropylene-ethylene copolymer, Cello Pack, Buffalo, N.Y.) as barrier material, first by folding the barrier material on itself to form the bottom28of the pouch assembly row111, and then dividing the pouch into twelve compartments12by heat-sealed seams54coextensive with the length of the compartments12. The top of the barrier material is sealed to one end of fitment414, which is made of Monprene, a thermoplastic elastomer (MP 1627 1.3, QST Inc., St. Albans, Vt.). This provides one-to-one communication between compartments12and the respective fitment cavities18. The plungers416are made of solid polypropylene with vacuum grease applied to the seal surface34. The diameter of the channels formed in fitment414, such as channels486,494,502,510for example, is kept at or smaller than 1 mm to minimize diffusion of fluid across cavities18. Vacuum port36has a diameter of 2 mm. The main passageway of the branched channels is etched on the front surface422of fitment414and covered by a portion of bottom layer24of the barrier material of row111of compartments12. Next, air is evacuated from the pouch assembly by use of a freeze-dryer (Virtis “Advantage”, Cardiner, N.Y.) at a vacuum of 7 Pa. The length of time of lyophilizing depends upon the volume of a reagent optionally provided therein. The volume of fitment cavities18is the adjusted to 100 μl by lowering the plungers416to the second position. The plungers416are then secured in position by comb470, as shown inFIGS. 5 and 6, for example. The pouch assembly416is then taken out of the vacuum chamber. Four hundred microliters each of water, mineral oil, and PCR mixture are separately prepared in 1 ml syringes. The PCR mixture comprises 0.2 mM DNTP, 1X IT buffer (Idaho Technology, Cat #1770, Salt Lake City, Utah), 0.04 U/μl Taq polymerase, 500 pg/μl human genomic DNA, 0.5 μM each of primers PC03 and PC04 (LightCycler manual, p27, 1997, Idaho Technology), and 3X SYBR® Green I dye (Molecular Probes, Eugene, Oreg.). Cannulas attached to each syringe are used to puncture seals through sample entry ports42, and the liquids from each syringe are individually withdrawn by force of vacuum into three fitment cavities18that are in communication with channels486,494. After the nine cavities are completely filled with liquid, the comb470is disengaged, and plungers416are twisted to seal access to the respective channel and sample entry port, and then lowered to the third bottom position within cavity18to dispense the liquid into compartments12. The microliter volume of liquid dispensed into each compartment12averaged 95.5±4.22. No appreciable difference was noted between the three samples, even though mineral oil has roughly 100 times higher viscosity than water or the PCR mixture. The pouch assembly was further inserted into an air thermal cycler (RapidCycler, Idaho Technology) with a modified lid so as to prevent escape of hot air from the chamber, and was exposed to 45 cycles of heating and cooling according to the referenced protocol (LightCycler manual, p31). After thermal cycling, the pouch assembly was placed on a UV transilluminater. The three compartments that contained the PCR mixture, but not those that contained mineral oil or water, were found to have 3 to 4 times higher fluorescence intensity than before thermal cycling, indicating successful amplification of a gene fragment. Amplification of DNA was further confirmed by gel electrophoresis.

In another example using the twelve-compartment pouch assembly410ofFIGS. 5-7B, PCR primers and dNTPs are dispensed into cavities18and freeze dried for 13 hours. A solution containing genomic DNA (500 pg/μl), buffer and Taq polymerase (0.04 U/μl) was prepared in a 1 ml syringe and dispensed into the pouch assembly416as described above by puncture of seal through port42by a cannula. The force of vacuum distributed the solution equally into three fitment cavities18. This operation was repeated four times so that all twelve cavities18were filled with solution. Then by twisting and lowering all of the plungers416, the samples were transferred into the collapsible compartments12. The pouch assembly416was exposed to thermal cycling as described above, and all twelve reactions successfully produced amplified DNA products.

In yet another example using the twelve-compartment pouch assembly ofFIGS. 5-7B, PCR primers (SQF and SQR) and a fluorescent probe (SQP1) specific forSalmonella(described in detail in U.S. Patent Application 2003/0022177 A1, herein incorporated by reference) and dNTPs are dispensed into each cavity18and freeze dried as described above. Buffered Taq polymerase solutions (0.04 U/μl) containing four levels of dilutions ofSalmonellagenomic DNA (10 pg/μl, 1 pg/μl, 0.1 pg/μl, and 0.01 pg/μl) were prepared in 1-ml syringes. These solutions were then each dispensed into the pouch assembly as described above by puncture of each of the seals46, and each solution distributed equally into three fitment cavities18by force of vacuum so that all twelve cavities were filled. Once the samples were transferred into the collapsible compartments12, the pouch was inserted into a thermal cycler which cycles the temperature of the samples by successive squeezing actions of movable heating elements, as shown inFIG. 9and described by WO 03/007677 A2, herein incorporated by reference. All twelve reactions successfully produced amplified DNA products as indicated by the fluorescence signal being above background by cycle number50, as shown inFIG. 8. The timing of fluorescence signal emerging above background inversely correlates to the initial concentration of target DNA.

Looking now toFIG. 9, a PCR apparatus610is provided for use in temperature controlled processes such as amplification of DNA through thermocycling and detecting and analyzing a reaction through fluorescence. Illustrative PCR apparatus610includes a thermocycling subassembly612and a fluorimeter subassembly614positioned generally below thermocycling subassembly612. In general, thermocycling subassembly612subjects a reaction mixture or fluid sample56to temperature cycling, or repeated rounds of heating and cooling. Thermocycling subassembly612includes a first pair of heaters616,618and a second pair of heaters620,622. Illustrative heaters616and620are movable heaters, whereas illustrative heaters618and622are stationary heaters. Movable heaters616,620operate to squeeze the row111of compartments12back and forth so that the fluid samples56within compartments12are moved between the two temperature zones provided by first and second pair of heaters616,618and620,622. It is within the scope of this disclosure to include a thermocycling subassembly having more than two temperature cycling zones.

Pneumatic bladders624,626of thermocycling subassembly612operate to move respective heaters616,620back and forth. Although illustrative pneumatic bladders are disclosed, it is within the scope of this disclosure to move heaters616,620through the use of any suitable pressure-based actuator such as hydraulics, spring rows, etc., for example. As shown inFIG. 9, heater616is moved to a closed position to squeeze the fluid sample56to a lower portion of compartment12between the second pair of heaters620,622to be heated to the temperature of heaters620,622. After an appropriate duration, heater616is moved to an opened position (not shown) and heater622is moved from the opened position to the closed position to squeeze the fluid sample56to an upper portion of compartment12and into full thermal contact with first pair of heaters616,618to be heated to the temperature of heaters616,618. Thermal cycling is accomplished by repeating these steps.

Although the invention has been described in detail with reference to preferred embodiments, variations and modifications exist within the scope and spirit of the invention.