Patent Publication Number: US-2023158492-A1

Title: Set of chambers containing reagents

Description:
The invention relates to a set of chambers containing reagents to be used in a system and a device for detecting a target analyte, in particular a target nucleic acid, for instance DNA or RNA, by way of isothermal nucleic acid amplification and fluorescence. 
     Nucleic acid amplification technologies are used to amplify the amount of a target nucleic acid in a sample in order detect such target nucleic acid in the sample. A known nucleic acid amplification technology is Polymerase Chain Reaction (PCR). Isothermal nucleic acid amplification technologies offer advantages over polymerase chain reaction (PCR) in that they do not require thermal cycling or sophisticated laboratory equipment. 
     Known isothermal nucleic acid amplification technologies are inter alia Recombinase Polymerase Amplification (RPA) and Strand Invasion Based Amplification (SIBA) and other methods know to persons skilled in the art. 
     Recombinase polymerase amplification (RPA), is a method to amplify the amount of a target analyte, in particular a nucleic acid such as DNA or RNA in a sample. For recombinase polymerase amplification three core enzymes are used: a recombinase, a single-stranded DNA-binding protein (SSB) and a strand-displacing polymerase. Recombinases can pair oligonucleotide primers with homologous sequences in duplex DNA. SSB binds to displaced strands of DNA and prevents the primers from being displaced. The strand-displacing polymerase begins DNA synthesis at sites where the primer has bound to the target DNA. Thus, if a target gene sequence is indeed present in the tested sample, an exponential DNA amplification reaction can be achieved to amplify a small amount of a target nucleic acid to detectable levels within minutes at temperatures between 37° C. and 42° C. 
     The three core RPA enzymes can be supplemented by further enzymes to provide extra functionality. Addition of exonuclease III allows the use of an exo probe for real-time, fluorescence detection. If a reverse transcriptase that works at 37 to 42° C. is added then RNA can be reverse transcribed and the cDNA produced amplified all in one step. 
     By adding a reverse transcriptase enzyme to an RPA reaction, it can detect RNA as well as DNA, without the need for a separate step to produce cDNA. It is an advantage of RPA that it is isothermal and thus only requires simple equipment. While RPA operates best at temperatures of 37-42° C. it still works at room temperature. 
     For detecting the presence of a targeted nucleic acid in a sample, fluorescence detection technique can be used. After the light source at specific wavelength illuminates on the targeted nucleic acids, the DNA-binding dyes or fluorescein-binding probes of the nucleic acids will react and enable fluorescent signals to be emitted. The fluorescent signal is an indication of the existence of the targeted nucleic acids. 
     It is an object of the invention facilitate the testing of samples by means of nucleic acid amplification technology. 
     According to the invention, a set of containers is provided that comprises a first container, in particular a lysis container that encloses at least a first chamber and a second container, in particular a test container that encloses at least a second chamber. 
     The lysis container may contain a liquid lysing fluid that causes lysing of the cells in a sample to thus release the nucleic acids (DNA or RNA). The lysing fluid may comprise an acid, e.g. HCl or a weak alkali, and a surface active agent. 
     The test container contains a mixture of chemicals that can cause an amplification of nucleic acid in a sample. Preferably, the mixture comprises a recombinase, a single-stranded DNA-binding protein (SSB) and strand-displacing polymerase that causes a recombinase polymerase amplification (RPA). The test container further preferably contains exonuclease III allows the use of an exo probe for real-time, fluorescence detection. The mixture may be provided in form of a dry pellet. 
     According to a first aspect of the invention the first container, the second container and the fluid transfer interface are configured to allow a limited amount of fluid being transferred from the first chamber into the second chamber. The fluid transfer interface initially is closed so that the first container can be handled as a separate, closed unit. 
     In particular, a set of at least two distinct containers is provided that can be combined to form a single, fluid tight assembly, wherein a first container encloses at least a first chamber containing a first set of chemicals and/or agents and wherein a second container comprises at least a second chamber containing a second set of chemicals and/or agents that are at least in part distinct from the chemicals and/or agents of the first set of chemicals and/or agents. The first container and/or the second container comprises a septum that initially is fluid tight and that can be altered to form a passageway between the first container and the second container when the first container and the second container are combined to form a single, fluid tight assembly, in order to allow a controlled transfer of fluid from the first container to the second container. 
     Preferably, the first container and the second container are arranged in a housing that comprises means for altering the distance between the first container and the second container so as to selectively provide a first relative distance between the first container and the second container wherein the first chamber and the second chamber (enclosed by the respective first container and second container are fluidly separated and a second relatively closer distance between the first container and the second container wherein the first chamber and the second chamber are in fluid communication. 
     The means for altering the distance between the first container and the second container preferably comprise a helical member, in particular a helical grove that is configured to translate a rotation into an axial movement of one container with respect to the other container. 
     In a preferred embodiment, the first container is held in the housing by a releasable snap fit connection that is released if a force exerted by the second container on the first container exceeds a predetermined threshold. 
     A second aspect of the invention is a set of at least two distinct containers that can be combined to form a single, fluid tight assembly. The first chamber that is enclosed by the first container comprises a first set of chemicals and/or agents. Preferably, the first chamber comprises a fluid for lysing a sample. Accordingly, the first container preferably is a lysis container and the first chamber is a lysis chamber. The first chamber is closed prior to use. The second chamber that is enclosed by the second container comprises a second set of chemicals and/or agents that are at least in part distinct from the chemicals and/or agents of the first set of chemicals in the first chamber. Preferably, the second set of chemicals comprises chemicals required for testing for an analyte and in particular to amplify nucleic acids in a sample. Accordingly, the second container preferably is a test container enclosing a test chamber. The first container comprises an interface, in particular a lid, that can be opened when the first container and the second container are combined to form a single, fluid tight assembly, in order to allow contents of the first chamber to enter the second chamber. 
     The second aspect of the invention can implemented in combination with the first aspect of the invention or independently from the first aspect of the invention. 
     In any embodiment, the test container may comprise more than one, e.g, two, three or four separate test chambers. In each test chamber a different mixture of chemicals can be contained thus allowing for testing for different target analytes simultaneously. 
     Preferably each container is an integral unibody. In an alternative embodiment, the assembly of the first container and the second container is an integral unibody. 
     Preferably, the containers can be connected by means of a fluid tight snap fit connection to thus form a single, fluid tight assembly. As an alternative to a snap fit connection, a screw lock connection similar to a Luer-lock can be provided for tightly connecting the containers. Another alternative is a press fit connection between the containers. 
     The advantage of a fluid tight assembly of at least two containers is that the assembly can be disposed easily without the risk of contamination because the contents of the assembly is tightly secured within the assembly. 
     Preferably, the second container has transparent walls that allow excitation and detection of luminescence and in particular fluorescence. 
     An assembly comprising two initially separate containers and a fluorescence detection device with a receptacle that can receive the assembly for luminescence detection makes it possible to perform a testing method that comprises two consecutive chemical or biochemical method steps—for instance lysis and amplification—and a luminescence testing step in a simple, clean and safe manner that minimizes the risk of contamination and infection while providing easy handling. The fluorescence detection device can be re-used because in use it is not contacted by the sample or any agents or chemicals since these are tightly enclosed in the assembly of containers. The assembly of containers and its contents can be safely disposed after use because the contents is reliably kept within the interior of the assembly. 
     In a preferred embodiment, the system also comprises a piston for transferring fluid from the first chamber to the second chamber. The piston is configured to fit into the first chamber. Preferably, the travel of the piston is limited and corresponds to the amount of fluid to be transferred into the second chamber. The piston can comprise a lid for closing the lysis container and the test container. 
     The combination of the lysis container and piston is arranged such that in the event of the piston moving into the lysis container pressure release is possible through venting. The venting means are configured to prevent content of either the lysis container or the test container or both from escaping out of the respective chamber or chambers. 
     The first containers lid that allows fluid transfer from the first chamber to the second chamber, when the first container and the second container are connected, can be configured to be opened by connecting the first container and the second container. 
     According to a further aspect, a set of containers is provided, comprising at least one test container and a sample container that is configured to for collecting a fluid sample. In particular, the test container is configured for collecting a sample by way of gargling or mouth washing. By such embodiment, sampling by means of a swab is replaced by a test using mouthwash or gargling. This embodiment follows the basic concept as disclosed herein before featuring modified parts so that a simple testing can be achieved by mouthwashing and/or gargling. The reliability of the samples and subsequent test process is improved and the inconveniences of sample taking with a swab are avoided. 
     The set of containers preferably comprises a sample container, a lysis container and a test container. 
     Alternatively, the set of containers comprises a sample container that comprises a sample chamber and a compartment for a lysing fluid, said compartment initially being separated from the sample chamber. In such embodiment, no separate lysis container is needed. 
     In yet another embodiment, the sample chamber of the sample container can comprise a liquid that can be used for gargling and that can lyse a sample thus acquired. 
     The set of container is part of a system that further comprises a fluorescence detection device. 
     The system is configured to implement an isothermal amplification method such as RPA and SIBA and other methods, preferably isothermal methods. The amplification method is configured to be carried out in a temperature range between 25° C. and 47° C. In various embodiments, initial heating may be applied. Other embodiments implement continuous heating. There are also embodiments without any external heating. in one or more of the method steps. 
     The fluorescence detection device comprises
         a detection chamber configured to receive the second container, i.e. the test container or the contents of the test container,   a light source,   an optical sensor   an energy supply   a wireless data interface and   a controller.       

     The controller can be a microcontroller and/or a state machine. 
     The fluorescence detection device may further comprise heating means that allow heating of a test container that is inserted in the fluorescence detection device. 
     Preferably, the detection device and the second container comprise mutually engaging features that prevent a relative rotation between the detection device and the second container. 
     The detection chamber and the receptacle preferably are arranged in a truncated cone shaped protrusion of a housing of the detection device. The shape of sad preferably matches an inner shape of a flared section of a housing of a fluid tight assembly that includes at least the first and the second container. 
     The system can either be a point of care (POC) system wherein the fluorescence detection device is arranged at a point of care, for instance in a medical doctors office. Alternatively, the system may be a personal system wherein the fluorescence detection device is self-contained and mobile, in particular pocketable. 
    
    
     
       The invention shall now further be illustrated by way of an example and with a reference to the figures. Of the figures, 
         FIG.  1   : shows a detection system for detecting a target analyte by way of isothermal nucleic acid amplification and fluorescence; 
         FIG.  2   : shows a first embodiment of a set of containers to be used with the detection system of  FIG.  1   ; 
         FIG.  3   a,b   : show a piercing fluid transfer interface to establish a fluid connection between the lysis chamber and the test chamber; 
         FIG.  4   : shows schematic diagrams of a detection device of the system from  FIG.  1   ; 
         FIG.  5   a - c   : show a second embodiment of a set of containers to be used with the detection system of  FIG.  1   ; 
         FIG.  6   a,b   : show a third embodiment of a set of containers to be used with the detection system of  FIG.  1   ; 
         FIG.  7   : shows a fourth embodiment of a set of containers to be used with the detection system of  FIG.  1   ; 
         FIG.  8   a - c   : show a fifth embodiment of a set of containers to be used with the detection system of  FIG.  1   ; 
         FIG.  9   : shows a sixth embodiment of a set of containers to be used with the detection system of  FIG.  1   ; 
         FIG.  10   : shows a seventh embodiment of a set of containers to be used with the detection system of  FIG.  1   : 
         FIG.  11   : details of the seventh embodiment of  FIG.  10   ; 
         FIG.  12   a, b   : show an eighth embodiment of a set of containers to be used with the detection system of  FIG.  1   ; and 
         FIG.  13   : shows a lysis chamber with cutting means for cutting a test swab; 
         FIG.  14   a   : is an isometric view of a further embodiment of a set of containers forming a fluid tight assembly to be used with the detection system of  FIG.  1    or the detection device of  FIGS.  18  and  19   ; 
         FIG.  14   b   : is an isometric view of the fluid tight assembly from  FIG.  14   a    with an open lid; 
         FIG.  15   a   : is a front elevation view of the fluid tight assembly from  FIG.  14   ; 
         FIG.  15   b   : is side elevation view of the fluid tight assembly from  FIG.  14   ; 
         FIG.  16   a   : is a top view of the fluid tight assembly from  FIG.  14   ; 
         FIG.  16   b   : is a bottom view of the fluid tight assembly from  FIG.  14   ; 
         FIG.  17   a   : is a cross sectional view of the fluid tight assembly from  FIG.  15     a;    
         FIG.  17   b    is a cross sectional view of the fluid tight assembly from  FIG.  15     b;    
         FIG.  18   : is a cross sectional view of a testing device adapted to match the fluid tight assembly from  FIGS.  14  to  17   ; and 
         FIG.  19   a   : is an exploded side view of the testing device of  FIG.  18   ; 
         FIG.  19   b   : is an exploded isometric view of the testing device of  FIG.  18   ; and 
         FIG.  20   a - c   : show an alternative embodiment having a third container for collecting a fluid sample. 
     
    
    
     The system  10  for detecting a target analyte comprises a fluorescence detection device  16  and a set of two containers  12  and  14 . A first container  12  is a lysis container that encloses a first chamber  13  comprising a fluid and/or an agent for lysing a sample. The second container  14  is a test container  14  that encloses a second chamber  15  comprising reagents, for instance enzymes for amplifying nucleic acids in a sample that was lysed in the lysis container  12 ; cf.  FIG.  1   . 
     The set of two containers  12  and  14  further comprises means that allow a limited transfer of fluid from the first chamber  13  into the second chamber  15 . In the embodiment shown in  FIGS.  1  and  2   , the means for limited transfer of fluid from the first container  12  to the second container  14  comprise a piston  18 . 
     The lysis container  12  contains a lysing fluid that causes lysing of cells or viruses in a sample to be tested. By way of lysing, nucleic acids such as DNA or RNA are released by way of breaking down the cells or viruses in the sample to be tested. Lysing can be achieved by a lysing fluid that comprises an acid such as hydrochloric acid or a weak alkali. 
     The lysis container  12  has a lid  20  so lysis container  12  can be opened and a sample to be tested can be entered in the lysis container  12 ; cf.  FIG.  2   . The contents of the lysis container  12  is about 100 μl. 
     Lid  20  of the lysis container  12  preferably is a membrane  20  that can be pierced by a cotton swab with a sample. The membrane  20  can be a composite film, for instance made from plastic sheeted aluminum. Alternatively, the membrane  20  can be an integral part of the lysis container  12 , in particular a thin septum formed from the material of the lysis container walls. Lid  20  can alternatively be a cap that can be opened and closed. 
     The test container  14  encloses the second chamber  15  comprising a mixture of enzymes that are needed for a recombinase polymerase amplification (RPA). Preferably, the mixture is provided in the form of a dry pellet  21  that is contained in the second chamber. 
     The test container  14  preferably is a cuvette that can be inserted in a receptacle  22  of the fluorescence detection device  16 . The receptacle  22  is part of a detection chamber  42  of the fluorescence detection device  16 . 
     The test container  14  is dimensioned to allow inserting the lysis container  12  into the test container  14  (or vice versa) in a tight and defined manner. An abutment  24  limits the depth of insertion. Once the lysis container  12  is fully inserted into the test container  14 , the piston  18  can be used to transfer the fluid from the lysis container into the test container  14  and allowing the recombinase polymerase amplification to work in the closed test container  14 . To allow the transfer of the contents of the lysis container  12  into the test container  14 , a further thin lid  26  is arranged as a septum at the bottom of the lysis container  12 . The thin lid  26  is dimensioned to break under the fluid pressure caused by the piston. Alternatively, the piston  18  and the thin lid  26  can be designed so that a tip (not shown) of the piston  18  can pierce the thin lid  26 . 
     The test container  14  may also be closed by a septum, for instance a thin pierceable lid  28  or  28 ′. The pierceable lid  28  maybe arranged at the otherwise open end of a guiding sleeve  37  of test chamber  37  or close to the level of the abutment  24  (lid  28 ). 
     The thin lids  26  and  28  can be a composite film, for instance made from plastic sheeted aluminum. Alternatively, the thin lid  26  can be an integral part of the lysis container  12 , in particular a thin septum formed from the material of the lysis containers  12  walls. Further preferred materials for the thin lids  26  and  28  are silicone or a thermoplastic elastomer (TPE). 
     In order to create a fluid interface and establish a fluid connection between lysis container  12  and test container  14 , a piercing fluid transfer interface member  27  is provided; see  FIG.  3   . The piercing fluid transfer interface member  27  comprises a fluid passageway  29  formed in tube-shaped protrusions  31  and  33  that extend centrally from a support disc  35  in opposite directions. Each tube-shaped protrusion  31  and  33  has a pointed tip that can pierce lid  26  and  28 , respectively. The tube-shaped protrusions  31  and  33  maybe formed from a metal tube that extends through the support disc  35 . Alternatively, tube-shaped protrusions  31  and  33  maybe integrally formed with the support disc  35  from a plastic material. 
     Lysis container  12 , test container  14  and piercing fluid transfer interface member  27  are held and guided in the guiding sleeve  37 . The fluid passageway  29  extends centrally along a common longitudinal axis of the lysis container  12 , the test container  14  and the guiding sleeve  37 . Piercing fluid transfer interface member  27  could be fixed to guiding sleeve  37  or even an integral part of guiding sleeve  35  so that lysis chamber  12  and test chamber  14  could be inserted into guiding sleeve  35  until respective lid  26  or  28  is pierced. However, it is preferred if piercing fluid transfer interface member  27  can slide within guiding sleeve  37  because this would provide that both lids  26  and  28  are pierced simultaneously. This is, because a sliding piercing fluid transfer interface member  27  would be pushed by either lid  26  or  28  until piercing fluid transfer interface member  27  abuts the respective other lid  28  or  26 . Only when sliding fluid transfer interface member  27  touches both lids  26  and  28 , piercing of the lids would occur. 
     Test container  14  and guiding sleeve  37  could form an integral unibody similar to what is shown in  FIG.  2   . If the test container  14  and the guiding sleeve  37  are firmly connected so as to form an integral part, it is mandatory that the fluid transfer interface member  27  is slidingly arranged within guiding sleeve  37 .  FIG.  3 B  shows an enlarged detail from  FIG.  3 B  in semi-perspective view. 
     The lysis container  12 , the test container  14  and the piston  18  are configured to engage in a fluid-tight manner when fully inserted. Such fluid tight engagement can be achieved by means of a snap fit connection wherein an annular protrusion  30  of one of the lysis container  12  and the test container  14  engages in an annular groove of the respective other container. Likewise, an annular protrusion  32  of one of the piston  18  and the lysis container  12  engages in an annular groove of the respective other part. Annular protrusions  30  and  32  act as sealings and may be integrally formed with the rest of the respective container or piston. 
     As an alternative to a snap fit connection, a screw lock connection similar to a Luer-lock can be provided for tightly connecting the lysis container  12  and the test container  14 . 
     To allow insertion of the lysis container  12  into the test container  14  and of the piston  18  into the lysis container  12 , venting means (not shown) are provided. 
     Once fully engaged, the test container  14 , the lysis container  12  and the piston  18  form a tight assembly  34  that can be handled as a single, fluid tight unit. 
     Walls  36  of the test container  14  are transparent so as to allow light to enter the test container  14  and to exit the test container  14 . The transparent walls  36  of the test container  14  make it possible to expose the content of test container  14  to exiting light that can cause a luminescence. In case, the content of the test container  14  is luminescent luminescence of the sample in the test container  14  can be detected through the transparent walls  36  of the test container  14 . 
     The pellet  21  that contains the mixture of enzymes for recombinase polymerase amplification preferably comprises a recombinase, a single-stranded DNA-binding protein (SSB) and a strand-displacing polymerase, exonuclease III and in case RNA is to be detected, a reverse transcriptase. 
     In use, a sample to be tested first is filled into lysis container  12 . After lysing the cells in the sample to be tested, the entire content of lysis container  12  is transferred into the test container  14  by means of the piston  18  so that a recombinase polymerase amplification can occur in the test container  14 . 
     Once the recombinase polymerase amplification has occurred in the test container  14 —typically between 10 to 15 minutes after filling in the content of the lysis container  12  into the second chamber that is enclosed by the test container  14 —the test container  14  or alternatively only its content can be entered into the fluorescence detection device  18 . 
     In the illustrated, preferred embodiment, the entire assembly  34  is inserted in a receptacle  22  of the fluorescence detection device  16 ; cf.  FIG.  4   . For handling of the assembly  34 , a grip  38  is provided at a proximal end of the piston  18 . 
     In order to prevent external light, for instance stray light, from entering into the receptacle once the assembly  34  is fully inserted in the receptacle  22 , a collar  40  is provided that forms a lid for the receptacle  22 . 
     The fluorescence detection device  16  comprises a detection chamber  42  that is configured to receive the test container  14  or the contents of the test container  14 . In the illustrated, preferred embodiment, the receptacle  22  is part of the detection chamber  42 . Within the detection chamber  42  or adjacent to the detection chamber  42 , a light source  44  and an optical sensor  46  are arranged. The light source  44  is configured to illuminate the contents of the detection chamber  42  with a light that can cause luminescence in a sample to be tested during and after the sample has undergone recombinase polymerase amplification. The optical sensor  46  is arranged and configured to detect luminescence in the detection chamber  42  in case luminescence occurs. 
     The detection chamber  42  is arranged in a detection chamber housing  70  that has outer dimension smaller than 10 cm by 10 cm by 4 cm. Preferably the volume of the entire fluorescence detection device  16  is smaller than 200 cm 2 , and even more preferred smaller than 100 cm 2 . In a preferred embodiment, the longest outer dimension is a least twice as long as the shortest outer dimension. 
     The fluorescence detection device  16 ′ may provide a second receptacle wherein the lysis container  12  can be placed prior to the connection with the test container  14 . The second receptacle can also host the test container  14  and the lysis container  12  prior to use. 
     With respect to the lysis container  12  and the test container  14  it is an object to only transfer a limited amount of fluid from the lysis container  12  to the test container  14 . Preferably, the amount to be transferred from the lysis container  12  into the test container  14  is reproducible within a range of tolerance of ±20% of the transferred volume. For instance, it is an object to transfer about 50 μl from the lysis container  12  into test container  14 , while the total amount of liquid in lysis container  12  is larger, for instance more than 100 μl. 
     In  FIG.  2   , a level  80  of fluid is shown. Piston  18  is configured to only displace about 50 μl of the fluid in lysis container  12 . This is achieved by a piston  18  that is shorter than the length of the first chamber that is enclosed by the lysis container  12  so that bottom  82  of piston  18  will not reach the bottom  84  of the first chamber even if the piston  18  is fully introduced into the lysis container  12 . Thus, only a reproducible share of the fluid in lysis container  12  is transferred into the test container  14 . The transferred share of fluid may for example correspond to 50 μl of fluid. 
     An implementation of an assembly  34 ′ similar to the assembly  34  of  FIG.  2    is shown in  FIGS.  5   a, b  and  c   . Test container  14 ′ can be connected to lysis container  12 ′ by a snap fit connection. The cylindrical side wall of the lysis container  12  extends beyond a bottom  84 ′ and thus forms an extension  88 . Test container  14 ′ has an annular flange  90  that allows for a snap fit connection with the lysis container  12 ′ when the flange  90  engages with the extension  88 ; see  FIG.  5     b.    
     Test container  14 ′ is closed by a lid  28 ′ that is sealingly connected to the annular flange  90  of test container  14 . Lid  28 ′ preferably is made from an annular composite film. Alternatively, the thin lid  28 ′ can be an integral part of the test container  14 , in particular a thin septum formed from the material of the test containers  14  walls. 
     In the center of bottom  84 ′ of lysis container  12 ′, a pointed central tip  92  is arranged and a valve  94  extends through the central pointed tip  92 . The central pointed tip  92  maybe made from metal or from plastic material. 
     A piston  18 ′ with limited travel is integrated in a screw-on cap  96 . Screw-on cap  96  that can be screwed on lysis container  12 ′. Accordingly, a swab with a sample can be inserted into the first chamber that is enclosed by a lysis container  12 ′ and thereafter, lysis container  12 ′ can be closed by screwing on the screw-on cap  96 . Once lysis chamber  12 ′ is closed with the screw-on cap  96 , lysis container  12 ′ can be connected to test container  14 ′. When test container  14 ′ snaps into a lysis container  12 ′, the pointed tip  92  at the center of bottom  88 ′ of lysis container  12 ′ pierces into lid  28 ′. Then, piston  18 ′ can be pushed to thus press a limit amount of fluid from the first chamber enclosed by the lysis container  12 ′ into the second chamber that is enclosed by the test container  14 ′. 
     In yet another embodiment as shown in  FIGS.  6   a  and  b   , lysis container  12 ″ having a bottom  88 ″ with a pointed central tip  92  and a valve  94  arranged therein can be inserted into a housing  98 . Lysis chamber  12 ″ as an outer diameter that fits into a lysis container receptacle  100  that is formed by a test container extension  102 . Test container extension  102  is provided with a threating  104  on the outside of the test container extension  102 . 
     The housing  98  is provided with an inner threating  106  that fits to the outer threating  104  of the test container  14 ″. 
     When the lysis chamber  12 ″ is inserted into the housing  98 , housing  98  can be screwed on the test container  14 ″ to thus press the lysis container  12 ″ into the lysis container receptacle  100  of the test container  14 ″. Once the pointed central tip  92  at the bottom  88 ′ of lysis container  12 ″ pierces the lid  28 ′ that initially closed the second chamber in the test container  14 ″, fluid can be transferred from the lysis container  12 ″ into the test container  14 ″ by further compressing the lysis container  12 ″. Further rotating of housing  98  causes further compression of the lysis container  12 ″ until an annular opening of the lysis container  14 ″ overcomes an annular protrusion  114  on a central extension  116  of housing  98  extending into the lysis container  12 ″. Compression of the lysis container  12 ″ is thus limited by the force needed for the annular opening of the lysis container  14 ″ to overcome the annular protrusion  114 . 
     Prior to piercing lid  28 ″, a swab with a sample can be inserted into lysis container  12 ″ to have the sample lysed by the lysing fluid in the lysis container  12 ″. A central opening  108  in the central extension  116  of the housing  98  is provided that allows insertion of a swab into the first chamber that is enclosed by the lysis container  12 ″. The central opening  108  can be closed by a push-on cap  110 . In order to prevent the pointed tip  92 ″ from prematurely piercing the lid  28 ″ of test container  14 ″, a helical spring  112  can be provided, that pushes the lysis container  12 ″ away from the test container  14 ″. The spring force of spring  112  is overcome by turning housing  98  and thus screwing housing  98  further onto the test container  14 ″. 
     Instead of arranging the central pointed tip on the bottom of lysis container  12 ″, a central pointed tip could be arranged at the top of the test container instead of lid  28 ″. In such embodiment, a pierceable lid would be arranged at the bottom of the lysis container. 
     An alternative approach is provided by an assembly  34 ′″ as is illustrated in  FIG.  7   . The assembly  34 ′ also comprises a lysis container  12 ′″ and a test container  14 ′″. At the bottom  84 ′″ of lysis container  12 ′″ an elastic valve  86  is arranged that opens, when the test container  14 ′″ is inserted into an extension  88 ′″ of lysis container  12 ′″. 
     Initially, test container  14 ′″ is closed by a lid  28 ′″ that is made from an aluminum composite film. The chamber enclosed by test container is evacuated so that a vacuum exists within the test container  14 ′. The lid  28 ′″ of test container  14 ′″ is sealingly connected to an annular flange  90 ′″ of test container  14 ′″. 
     When the test container  14 ′″ is inserted into the extension  88 ′″ of the lysis container  12 ′″, the annular flange  90 ′″ will abut an annular, convex bulge  120  of valve  86  at the bottom  84 ″″ of the lysis container  12 ′″. This will cause a pointed center part  92 ′″ of valve  86  to pierce into the aluminum composite film  28 ′″ covering the test container  14 ′″. At the same time, a central hole  94 ′″ in the pointed central tip  92 ′″ of valve  86  will let a definite amount of fluid flow from the lysis container  12 ′″ into the test container  14 ″″. This is because initially, a vacuum exists within the test container  14 ′″. Elastic forces of the elastic valve  86  will keep a central opening  94 ′″ in the pointed central tip  92 ′″ of valve  86  closed until the annular bulge  120  of valve  86  is compressed in an axial direction of the lysis container  12 ′″ due to the abutting annular flange  90 ′″ of the test container  14 ′″. 
     In  FIGS.  8   a, b  and  c    a fluid transfer interface of yet another embodiment of an assembly comprising a lysis container  12 ″″ and a test container  14 ″″ is illustrated. At the interface between the lysis container  12 ″″ and the test container  14 ″″ a portioner  130  is arranged. Portioner  130  comprises a base member  132  having an annular groove  134  facing towards the first chamber that is enclosed by the lysis container  12 ″″. The annular groove  134  extends about 3400 to 350°. At one end of the annular groove  134  a through-hole  136  is provided that can be brought in alignment with an opening  138  being an fluid communication with the second chamber that is enclosed by the test container  14 ″″. A slide  140  is provided that can rotate with respect to the base member  132  to thus displace fluid in the annular groove  134 . 
       FIGS.  8 A to  8 C  illustrate the operation of the portioner  130 . In the initial position, an annular segment shaped opening  142  in the bottom  84 ″″ of the lysis container  12 ″″ is in fluid communication to the annular groove  134  so that fluid from the first chamber that is enclosed by the lysis container  12 ″″ can flow into the annular groove  134 . Rotating the lysis container  12 ″″ with respect to the portioner  130  closes the fluid passage between the first chamber and the annular groove  134 ; see  FIG.  8 B . The annular segment shaped opening  142  has moved due to rotation so that the annular groove  134  in the portioner  130  is closed. 
     An edge  144  of the annular segment shaped opening  142  engages an attachment  144  of slide  140  so that the slide  140  rotates together with the lysis container  12 ″″. Rotation of slide  140  causes also the base member  132  of portioner  130  to rotate because the fluid in the annular groove  134  cannot be compressed. Rotation of the base member  132  of portioner  130  occurs, until the through-hole  136  in the bottom of the annular groove  134  is aligned with the opening  138  of the test container  14 ″″ that is in fluid communication with the second chamber that is enclosed by the test container  14 ″″; cf.  FIG.  8 C . 
     A radial extension  146  of the test container  14 ″″ can engage with a groove in the receptacle  22  of the detection chamber  42  of the fluorescence detection device  16 . The radial extension  146  prevents the test container  14 ″″ from rotating. 
     In yet another embodiment, as shown in  FIGS.  9  to  11   , the lysis container  12 ′″″ may comprise the first chamber  150  and a third chamber  152  and a movable separation member  154 . The movable separation member  154  is configured to be moved between a first state and a second state. In the first state, the movable separation member  154  allows fluid to pass from the first chamber  150  to the third chamber  152 . In the second state, movable separation member  154  blocks fluid from passing from the first chamber  150  to the third chamber  152 . The movable separation member  154  preferably is an integral wall part of lysis container  12 ′″″ that is connected to a wall of lysis container  12 ′″″ by a breaking score line  156 . Breaking of the score line  156  occurs, when the lysis container  12 ′″″ is connected with the test container  14 ′″″ and causes the movable separation member  154  from moving from the first state to the second state. The third chamber  154  that is enclosed by the lysis container  12 ′″″ encloses a volume that corresponds to the volume of liquid that is to be transferred from the first chamber in the lysis container  12 ′″″ to the second chamber in the test container  14 ′″″. 
     When the lysis container  12 ′″″ and the test container  14 ′″″ are connected, a fluid passage is established between the third chamber  152  in the lysis container  12 ′″″ and the second chamber in the test container  14 ′″″ thus allowing a definite amount of fluid flowing from the third chamber  152  into the second chamber. 
     In yet another embodiment of assembly  34 ″″″ comprising a lysis container  12 ″″″ and a test container  14 ″″″ as shown in  FIGS.  12   a  and  b   , lysis container  12 ″″″ encloses a first chamber  150 ″″″ and a third chamber  152 ″″″. The third chamber  152 ″″″ is defined by a cylindrical side wall  160  and by the bottom  84 ″″″ of the lysis container  12 ″″″. The end of the third chamber  151 ″″″ that faces away from the bottom  84 ′″ is open so that the first chamber  150 ″″″ and the third chamber  152 ″″″ are not separated. The inner diameter of the third chamber  152 ″″″ fits to an outer diameter of a piston  18 ″″″ that thus can be inserted into the third chamber  152 ″″″. Once a bottom  82 ″″″ of piston  18 ″″″ engages with the side wall  160  of the third chamber  152 ″″″, the first chamber  150 ″″″ and the third  152 ″″″ are fluidly separated from one another. The volume enclosed by the side wall  160  is about 50 μl to 70 μl. 
     In order to transfer fluid from the lysis container  12 ″″″ to the test container  14 ″″″, test container  14 ″″″ can be attached to the bottom  84 ″″″ of the lysis container  12 ″″″ and will be held by an extension  88 ″″″ of the lysis container  12 ″″″. Once attached, a pointed central tip  92 ″″″ pierces into a lid  28 ″″″ of the test container  14 ″″″. Further, a venting tip  162  pierces into the bottom  84 ″″″ of the lysis chamber  12 ″″″ outside of the third chamber  152 ″″″. This is shown in  FIG.  12 B . 
     In order to transfer fluid from the lysis chamber  12 ″″″ into the test chamber  14 ″″″, the piston  18 ″″″ is pushed into the lysis chamber  12 ″″″ and will not cause any fluid transfer from the lysis container  12 ″″″ to the test container  14 ″″″ until the bottom  82 ″″″ of the piston  18 ″″″ reaches the side wall  160  of the third chamber  152 ″″″. Then, the contents of the third chamber  152 ″″″ is pressed through essential opening in the central pointed tip  92 ″″″ into the second chamber enclosed by the test container  14 ″″″. Venting of the second chamber enclosed by the test container  14 ″″″ can occur via venting tip  162 . 
     For guiding a piston  18 ″″″, guiding ribs  164  are provided, that extend in the longitudinal direction of the lysis container  12 ′″″. 
     Lysis container  12 ″″″ may provide a guiding extension  166  for guiding the piston  18 ″″″. A hole  168  in the extension  166  allows inserting a tip of a swab  170 . When the piston  18 ″″″ is pushed into the lysis container  12 ″″″ after while a swab  170  extends through the hole  168  in the wall of the extension  166 , the swab is cut in parts and the part comprising the genetic material to be lysed is kept in the lysis container  12 ″″″; see  FIG.  13   . Note that the embodiment of  FIG.  13    corresponds to the embodiment of  FIG.  12   . 
     In alternative embodiments, a cross slide for cutting the swap may be provided, the cross slide would have a cutting edge and would be arranged to move in a direction that is perpendicular to the longitudinal direction of the lysis container. Cross slide guiding elements may be provided on the lysis container for guiding the cross slide. Preferably, the cross slide is arranged close to an end of the lysis container that faces away from the test container. 
     Alternative embodiments of integral cutting means for cutting a swab can be provided with the other embodiments disclosed in this description to thus provide further preferred embodiments. 
     In yet another embodiment (not shown in the figures), the lysis container is one part but consists of a lower hollow and an upper plain cylinder. The test container plugs into the hollow part of the lysis container from the bottom. The lysis container covers the test container on the edges. When the lysis container is full, the hollow cylinder is filled and the plain cylinder is filled with 50 μl to 70 μl. Now the surface where the two containers meet needs to be opened and pushed to the edges, covering the liquids in the hollow cylinder but allowing the amount of 50 μl to 70 μl in the plain cylinder to mix with the pellets. The surface of the inner circle and the outer ring (of the hollow cylinder) must be equivalent The required amount (50 μl to 70 μl) is roughly the volume of a water-droplet. 
     In yet another embodiment (not shown in the figures), the lysis container is connected to the test container with an M-shaped tube. The “M” gets filled and when you turn it back around, liquid remains in the middle curvage. That amount can be calculated and used for further steps. 
     An embodiment similar to the embodiment in  FIG.  6    is shown in  FIGS.  14  to  17   . 
     Lysis chamber  120  and test chamber  140  are hold in a housing  980  that is comprised of two parts, a lower housing part  98 . 1 ° and an upper housing part  98 . 2 °. The lower housing part  98 . 1 ° has a flared side wall  180  that widens towards the bottom of housing  98 °. The flared side wall  180  is configured to fit to a truncated cone shaped protrusion  182  of the detection device  160  (see  FIGS.  18  and  19   ). 
     The top of housing  980  is closed by a cap  1100  that can swivel into an open position (see  FIG.  14   b   ) for selectively opening a central opening  184  for inserting a swab into the lysis chamber  130  of lysis container  12 °. 
     In the upper part of housing  980  the lysis container  120  is held by means of a releasable snap fit connection achieved by a outwardly extending circumferential collar  186  around an upper section of the lysis container outer wall and two inwardly extending circumferential rips  188  on the inner wall of an upper section of housing  98 °. The outwardly extending circumferential collar  186  of the lysis container  120  is held in a grove between the two inwardly extending circumferential rips  188  on the inner wall of housing  98 °. The snap fit connection between the lysis chamber  120  and the housing  980  can be released by a force acting on the lysis chamber  120  in an axial direction that exceeds a predetermined threshold as provided by the design of the snap fit connection as provided inter alia by the matching shapes and the elastic properties of the materials and shapes. 
     In the bottom  840  of the lysis container  120  a hollow needle  200  is arranged. The needle  200  preferably is made from stainless steel. 
     Similar to the embodiment of  FIG.  6   , the lower part of the lysis container  120  including the lysis container bottom  840  is arranged to extend into a lysis container receptacle  1000  that is formed by a test container extension  1020  extending upwardly from an upper part of the test container  14 . The outer diameter of the lower part of the lysis container  120  corresponds to the inner diameter of the lysis container receptacle  100 °. 
     Similar to the embodiment of  FIG.  6   , a rotation of the housing  98  relative to the test container  14  causes a relative axial movement of the test container  14 . This is achieved by at least one helical grove  202  that is formed in the inner side of the wall of the housing  98 . In the illustrated embodiment, three helical groves  202  are provided. A radial protrusion  204  on the test container extension  1020  radially extends into the grove  202 . Thus, the grove  202  acts as a helical slotted link guide for the radial protrusion  204  on the test container extension  102 °. The helical grove  202  causes an axial movement of the test container  14  when the housing  980  is rotated while the test container  140  does not rotate. Accordingly, the helical grove  202  and the radial protrusion  204  are an alternative to the threading  104  of the embodiment of  FIG.  6   . 
     In order to prevent a rotation of the test container  140  when the housing  980  is rotated, a short longitudinal rip  206  extending in the radial direction from a wall of the test container  140  is provided. The rip  206  engages with a notch  208  of the receptacle  220  of the detection device  16 ° when the test container  140  of fluid tight assembly  340  is inserted on the receptacle  220  of the detection device  16 °. 
     Rotating of the housing  980  relative to the test container  140  causes an axial movement of the test container  140  towards the lysis container  12 °. This in turn causes the needle  200  to pierce the elastomeric septum  280  (i.e. the lid) of the test container  14 °. Once the needle  200  at the bottom  840  of lysis container  120  pierces the elastomeric septum  28 ° (separating wall acting as a lid for the test container that can be pierced by a needle) that initially closed the second chamber in the test container  14 °, fluid can be transferred from the lysis container  120  into the test container  14 °. 
     The axial movement of the test container  140  towards the lysis container  120  further causes a compression of the lysis container  120  and thus a transfer of fluid from the lysis chamber  130  into the test chamber  15 °. Further rotating of housing  980  causes further compression of the lysis container  120  until the axial forces on the lysis container  120  causing the compression of the lysis container  120  exceed the force needed to release the lysis container  120  from being held by the outwardly extending circumferential collar  186  of the lysis container  120  in the grove between the two inwardly extending circumferential rips  188  on the inner wall of housing  98 °. Once released, the lysis container  120  can freely move upwards (i.e. in the direction towards the central opening  184  at the top of housing  98 °) and the lysis container  120  is not further compressed. Thus, further transfer of fluid from the lysis chamber  13 ° into the test chamber  15 ° is stopped. Compression of the lysis container  12 ° is thus limited by the force needed for pushing the outwardly extending circumferential collar  186  of the lysis container  120  out of the grove between the two inwardly extending circumferential rips  188  on the inner wall of housing  98 °. 
     For using the fluid tight assembly  340  in combination with the detection device  16 °, first a swap with a sample to be tested is inserted in the lysis camber  130  via the central opening  184 . Next, cap  1100  is closed and lysis can occur in the lysis chamber  13 °. Once lysis has occurred, the fluid tight assembly  340  can be engaged with the detection device  160  by inserting the test container  140  into the receptacle  220  of the detection device  16 °. This is facilitated by the flared side wall  180  of the lower section of housing  98 °. 
     To further facilitate engaging the fluid tight assembly  340  with the detection device  160  in the right orientation that allows the short longitudinal rip  206  of the test container  140  to engage with the notch  208  of the receptacle  220  of the detection device  16 °, a palpable raised rip  210  is provided on the outer surface of the housing  98 °. The palpable raised rip  210  runs along a longitudinal direction of the housing  98 °. 
     A palpable protrusion  212  on the detection device  160  next to the truncated cone shaped protrusion  182  of the detection device  160  is a further palpable feature that helps orienting the palpable raised rip  210  and thus the fluid tight assembly  340  correctly. 
     Once engaged with the detection device  16 °, the housing  980  of the fluid tight assembly  340  can be rotated to cause the relative axial movement of the test container  140  relative to the lysis container  120  and the housing  98 °. Since immediately after engaging the fluid tight assembly  340  with the testing device  160  the test container  140  already is fully inserted into the receptacle  220  and thus the detection chamber  42 ° of the detection device  16 °, the housing  980  is axially spaced from the detection device  16 °. By rotating the housing  980  relative to the detection device  160  and thus the test container  14 °, the axial distance between the housing  980  and the detection device  160  is minimized until the housing  980  touches the detection device  160  and the needle  200  has pierced the septum  280  of the test container  14 °. As seen from outside, while rotating the housing  980  clockwise, the housing  980  moves downwardly thus approaching the detection device  16 °. 
     Once the housing  980  is rotated in its final position, a further effect of the truncated cone shaped protrusion  182  of the detection device  160  in combination with the flared side wall  180  of the housing  980  is an improved protection from straylight entering the detection chamber  420  of the detection device  16 °. 
     The detection chamber  420  of the fluorescence detection device  160  is configured to receive the test container  14 °. The receptacle  220  is part of the detection chamber  42 °. The light source  440  comprises two light emitting diodes that are arranged to illuminate the contents of the test chamber  150  in the detection chamber  420  with a light that can cause luminescence in a sample to be tested during and after the sample has undergone recombinase polymerase amplification. The optical sensor  460  is arranged and configured to detect luminescence in the detection chamber  420  in case luminescence occurs. 
     For controlling the light emitting diodes  440  and reading out and processing the signal provided by the light sensor  46 °, a controller  500  is provided that is operatively connected to a wireless data interface  520  comprising an NFC antenna. 
     Energy is supplied by a recharge battery  480  that can be charged via an USB-C port  214 . 
     Yet another embodiment of a fluid tight assembly is illustrated in  FIGS.  20   a ,  20   b  and  20   c   , a third container  220  is provided that serves for collecting a liquid sample instead of a sample collected by means of a swab. 
     The third container can be a separate sample container  220  that is used for taking a sample for instance by way of gargling or mouthwash. The sample container can be connected to a lysis container  12   vii  and a test container  14   vii . The lysis container  12   vii  and the test container  14   vii  can be configured as disclosed herein above. A combination of a lysis container and a test container is called reaction containers hereinafter. The reaction containers, i.e. the test container  14   vii  and the lysis container  12   vii  can form a single unitary unit while the sample container can be a separate component that can be attached to the reaction containers. 
     The sample container  220  comprises a sample chamber  222  that can be closed by a sample container lid  224 . 
     The sample chamber can be initially empty or it may contain a liquid such as pure water, a solution containing H 2 O 2  or the like that can be used for gargling or mouth washing. 
     The sample container  220  is used for sampling while lysing and the test reaction, for instance the amplification, is performed in the reaction containers, i.e. the lysis container  12   vii  and the test container  14   vii , respectively. 
     The correct dosing of the sample liquid is done through and during the connection of the two separate parts. 
     The containers can be connected, for example, by plugging them together or by a rotary movement, e.g. screwing, or a combination of both, for instance by way of a bayonet connection. Preferably, the containers are designed so that the movement for connecting the containers (plugging, screwing etc.) trigger a transfer of a defined amount of sample liquid to the reaction containers, thus triggering the detection reaction. 
     Preferably, in a first step, sample liquid is transferred from the sample container  220  into the lysis chamber in the lysis container  14   vii . 
     In second step, after lysing has occurred, liquid is further transferred to the test chamber of test container  14   vii . The amount of liquid transferred into the test chamber preferably is less than 100 μl, more preferred less than 50 μl. The amount depends and is adapted to the respective test. 
     In one embodiment, the sample container contains the liquid for sampling. A test person can take the liquid up for rinsing the mouth. Thereafter, the liquid comprising the sample is put back into the sample container. Then, the sample container is closed and can be connected with the reaction containers  12   vii  and  14   vii . 
     Typically, the sample liquid initially contained in the sample chamber is such that it can be spat into the mouth of test persons without danger and that it can collect the sample obtained by mouthwash. 
     According to a first embodiment, the sample liquid contained in the sample container can be a liquid that can lyse the viruses or bacteria so that a separate lysis step is no longer necessary. This could typically be realized by an oxidation reaction, e.g. by a 1-3% H 2 O 2  solution. In such embodiment, no separate lysing chamber is needed. 
     In yet another embodiment, no liquid is kept in the sample container for sampling, the test person rinses his mouth with cold tap water and spits this rinsing liquid into the sample container. The sample container also contains another liquid for the lysis of the sample. 
     The two liquids are only brought together after the sampling has taken place, i.e. the test person has spit the rinsing liquid into the sample container. In this embodiment, separate compartments are provided in the sample container to separate the lysing liquid from the sample liquid before mixing. By means of a mechanical, thermal or chemical process the two liquids can be treated in such a way that the liquids mix homogeneously after collecting the sample has occurred. 
     In both cases, the sample container and the reaction containers are connected to each other by plugging or twisting, e.g. by a bayonet fastener  226  as shown in  FIG.  20   b   ). In one embodiment, the arrangement of the lysis chamber and the sample container can be designed such that the sample container and lysis chamber are arranged on top of each other, as shown in  FIG.  14   . In alternative embodiments the containers are designed to be arranged side by side. In yet another embodiment, already lysed sample can be added to the sample container, thus avoiding a separated lysing step. 
     When connecting the containers, a mechanism is actuated which allows a transfer of a defined amount of liquid (of the sample) into the reaction containers. The reaction can then take place. Fluid transfer means  228 —as seen in the top view depicted in  FIG.  20   b   )—allow the transfer of fluid.