Reagent container and method for operating the container

Abstract

The present invention relates to a reagent container and a method for operating the container. The reagent container comprises a container body, a first sealing cover, a sealant and a second sealing cover. The reagent container has improved sealing performance.

Description

Technical Field

[0001] The present disclosure relates to the field of biological detection technology, and in particular to a reagent container and a method for operating the container. Background Art

[0002] Polymerase chain reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. This method can efficiently amplify selective gene fragments in vitro, thereby enabling the detection of target genes.

[0003] Since PCR products accumulate exponentially during the amplification process, when the PCR reaction is completed, the product concentration in the PCR tube is generally tens of millions of times higher than before the reaction or even higher. Once the PCR tube is not sealed tightly, such a high concentration of product will form an aerosol and leak, causing serious contamination, leading to abnormalities in subsequent PCR tests.

[0004] During the nucleic acid amplification operation, it is sometimes necessary to add or remove part of the reagent into or from the container. In this case, it is necessary to open the lid of the reagent container, add or remove the reagent, and then close the lid. Summary of the Invention

[0005] The inventors have discovered that existing reagent containers have the following problems:

[0006] (1) The container is sealed only by the mechanical seal formed between the container cover and the container wall, which is a single seal and there is a risk of loose sealing;

[0007] (2) Since high-concentration and volatile aerosols are generated during and after the nucleic acid amplification reaction, if such gases leak into the laboratory, they will cause laboratory contamination;

[0008] (3) When adding / removing reagents, the container cover needs to be opened, and the existing solution is difficult to realize automated operation.

[0009] (4) There is still a risk of reagent leakage when the container is left standing for use after use.

[0010] The present disclosure provides a reagent container, which has at least the following advantages:

[0011] (1) When adding / removing reagents, there is no need to open the container cover, which reduces the difficulty of operation, improves the operating efficiency, and facilitates automated operation;

[0012] (2) During the use of the container, in addition to the seal between the container cover and the container wall, an additional sealing measure is provided, reducing the risk of leakage of the reagent in the container;

[0013] (3) After the container is left to rest for use, in addition to the seal between the container cover and the container wall, an additional sealing measure is provided, reducing the risk of leakage of the reagent in the container.

[0014] In some aspects, the present disclosure provides a reagent container comprising:

[0015] Container body,

[0016] The container body includes a bottom and a side wall, and the bottom and the side wall define a cavity;

[0017] First sealing cover,

[0018] The first sealing cover is capable of sealingly cooperating with the side wall, thereby defining a sealed cavity together with the side wall and the bottom of the container body;

[0019] The first sealing cover is provided with a penetrable area;

[0020] sealants,

[0021] The sealant is positioned so as to be located within the sealed cavity when the first sealing cover, the sidewalls, and the bottom collectively define a sealed cavity;

[0022] The sealant is capable of undergoing a solid-liquid phase transition as environmental conditions change; and

[0023] The second sealing cover,

[0024] The second sealing cover can be in a separated state and a fitted state;

[0025] The separated state means that the second sealing cover does not form a sealed cavity with the container body;

[0026] The mating state refers to that the second sealing cover and the container body define a sealed cavity.

[0027] The advantages of the above reagent container are:

[0028] (1) By operating a penetrating mechanism (such as a pipette), the penetrable area on the first sealing cover can be penetrated, thereby adding reagents to the sealed cavity or removing reagents from the sealed cavity without the need for additional lid opening operation; this improves operating efficiency and facilitates automated operation;

[0029] (2) By changing the environmental conditions, the sealant is liquefied to form a liquid sealing layer, which is then combined with the seal formed by the second sealing cover and the container body to form a double sealing system, greatly reducing the risk of reagent leakage;

[0030] (3) After the container is used, the sealant forming the liquid sealing layer can be solidified to form a solid sealing layer. The solid sealing layer and the second sealing cover form a double sealing system, thereby greatly reducing the risk of reagent leakage.

[0031] In some embodiments, the mating state refers to the second sealing cover being sealingly mated with the side wall or the first sealing cover, thereby defining a sealed cavity with the container body.

[0032] In some embodiments, the penetrable region isolates the sealed cavity from the external environment. When the penetrable region is penetrated, the seal of the sealed cavity is broken and the cavity is connected to the external environment, thereby enabling the step of adding / removing reagents.

[0033] In some embodiments, the pierceable region may be located in the central region of the first sealing cover, which is convenient for piercing. The area of ​​the pierceable region may account for 1 to 100% of the cross-sectional area of ​​the first sealing cover.

[0034] In some embodiments, the penetrable region of the first sealing cover is a pierceable membrane.

[0035] In some embodiments, the penetrable region of the first sealing cover is a plastic film or a metal foil.

[0036] In some embodiments, when the second sealing cover is in the separated state, the second sealing cover is completely separated from the container body.

[0037] In some embodiments, after the penetrable area of ​​the first sealing cover is penetrated, the container body can be resealed by adjusting the second sealing cover to a mating state.

[0038] In some embodiments, when the second sealing cover is in the engaged state, the second sealing cover is sealed against the sidewall of the container body, and the second sealing cover and the container body define a second sealed cavity. In this case, the pierced first sealing cover can be sealed in the second sealed cavity.

[0039] In some embodiments, when the second sealing cover is in the mating state, the second sealing cover is sealed with the first container cover, and the second sealing cover, the first sealing cover, and the container body define a second sealed cavity. In this case, the penetrated penetrable area can be sealed in the second sealed cavity.

[0040] In some embodiments, when the first sealing cover, the sidewalls, and the bottom of the container body collectively define a sealed cavity, the top of the first sealing cover is lower than the top edge of the sidewalls. Thus, a reserved space is left between the top of the first sealing cover and the top edge of the sidewalls. After the penetrable area of ​​the first sealing cover is penetrated, the second sealing cover can be placed in the reserved space to re-form the sealed cavity with the container body.

[0041] In some embodiments, the first sealing cover is provided with a mating portion (e.g., a mating surface, a mating recess, a mating protrusion, or an annular mating portion, such as a groove, an annular wall, an annular step surface, etc.) that can be sealed with the second sealing cover, and correspondingly, the second sealing cover may also be provided with a mating portion (e.g., a mating surface, a mating recess, a mating protrusion, or an annular mating portion) that can be sealed with the first sealing cover. Thus, after the penetrable area on the first sealing cover is penetrated, the penetrated penetrable area can be sealed by sealingly engaging the second sealing cover with the first sealing cover, thereby achieving resealing of the sealed cavity.

[0042] In some embodiments, the first sealing cover is provided with a mating recess. The mating recess may have an upward opening and may include sidewalls and a bottom. The penetrable region is provided at the bottom of the mating recess. In this case, the outer edge of the second sealing cover seals against the inner sidewalls of the mating recess in a circumferentially sealed manner, thereby blocking the penetrated penetrable region and achieving resealing of the sealed cavity.

[0043] In some embodiments, the first sealing cover is provided with a mating protrusion, and the second sealing cover is correspondingly provided with a mating recess. The mating protrusion may protrude upward. The mating protrusion may include a mating protrusion sidewall and a top, and the penetrable area is provided at the top of the mating protrusion. In this case, the mating recess of the second sealing cover can seal the penetrated penetrable area by circumferentially sealing with the outer sidewall of the mating protrusion of the first sealing cover, thereby achieving resealing of the sealed cavity.

[0044] In some embodiments, when the nucleic acid amplification container is not in use, the sealant is in a solid state. When the container is in use, the environmental conditions can be modified as needed to liquefy the solid sealant, thereby forming a liquid sealing layer (liquid sealing layer) within the container body, thereby preventing the reagents and the aerogel formed by the reagents from overflowing the container during the reaction. After the reaction is completed, the environmental conditions can be modified as needed to solidify the liquid sealing layer, forming a solid seal (solid sealing layer).

[0045] In some embodiments, after all required reagents have been added to a nucleic acid amplification reaction, environmental conditions (such as temperature, light, or chemical environment) are modified to cause the sealant to change from a solid state to a liquid state and flow to the surface of the reagents, thereby forming a liquid seal on the surface of the preparation already in the container, preventing spillage of the preparation during subsequent reactions. After the nucleic acid amplification reaction is completed, environmental conditions are modified to cause the sealant to change from a liquid state to a solid state, forming a solid seal, preventing leakage of the preparation within the container after it has been left unused.

[0046] In some embodiments, the change in environmental conditions refers to a change in temperature, a change in light conditions, or a change in the chemical environment.

[0047] In some embodiments, the sealant is solid at a temperature below (including) T1 and liquid at a temperature above (including) T2, where T2 > T1. In this case, heating the container body liquefies the sealant, forming a liquid sealing layer within the tube. Cooling the container body solidifies the sealant in the liquid sealing layer, forming a solid cap.

[0048] In some embodiments, T1 = 20-40°C, such as 30-40°C.

[0049] In some embodiments, T2 = 45-95°C, such as 60-90°C, such as 70-80°C.

[0050] In some embodiments, heat radiation or light radiation may be applied from outside the container body to the sealant inside the container body by a temperature control device, a lighting device, or the like, thereby achieving control over the solid-liquid state of the sealant.

[0051] In some embodiments, the sealant has a density less than that of water.

[0052] In some embodiments, the sealant is insoluble in water.

[0053] In some embodiments, the sealant is affixed to the first sealing cap or within the container body.

[0054] In some embodiments, the density of the sealant after liquefaction is less than the density of the nucleic acid amplification reagent.

[0055] In some embodiments, the sealant is insoluble in the reagents for nucleic acid amplification.

[0056] In certain embodiments, the sealing reagent is a solid and can be converted into a liquid under predetermined conditions, and the density of the liquid is less than the density of the reagent for nucleic acid amplification, and the liquid is immiscible with the reagent for nucleic acid amplification (for example, it is immiscible with aqueous solutions and is non-volatile). In certain embodiments, the predetermined conditions are selected from heating, illumination and / or chemical treatment. For example, in certain embodiments, the sealing reagent is a solid and can be converted into a liquid under heating conditions, and the density of the liquid is less than the density of the reagent for nucleic acid amplification, and the liquid is immiscible with the reagent for nucleic acid amplification (for example, it is immiscible with aqueous solutions and is non-volatile).

[0057] As used herein, the term "immiscible" means that when two or more liquids are placed in the same container, they will not dissolve in each other, and when they come into contact, a dividing interface will spontaneously form, forming layers.

[0058] As used herein, the term "stratification" refers to the spontaneous formation of an upper and lower distributed structure when two or more immiscible liquids are placed in the same container due to their density difference, with the liquid with higher density sinking and the liquid with lower density floating to form stratification.

[0059] As used herein, the sealing reagent of the present invention is immiscible with nucleic acid amplification reagent in a liquid state, and its density is less than the density of nucleic acid amplification reagent.Therefore, when the sealing reagent of the present invention is in the same container as the nucleic acid amplification reagent in a liquid state, the two will form a layered structure, and the sealing reagent of the present invention will be located in the upper layer, isolating (that is, sealing) the nucleic acid amplification reagent (and other substances that may be contained therein, such as nucleic acid) located in the lower layer, both preventing the lower layer reagent (and other substances that may be contained therein, such as nucleic acid) from escaping to the external environment, and also preventing the substances in the external environment from entering and contaminating the lower layer. In the present application, it is preferred that nucleic acid molecules cannot be dissolved in the sealing reagent of the present invention. Thus, the sealing reagent of the present invention can better play an isolating role. In addition, it is understandable that any substance that can achieve this function can be used as the sealing reagent of the present application, and therefore, the sealing reagent of the present application is not limited to the substances listed in the disclosure.

[0060] In some embodiments, the sealant is a wax, such as paraffin wax or EVA wax (ethylene vinyl acetate copolymer).

[0061] In some embodiments, the sealant is secured to the sidewalls.

[0062] In some embodiments, the sealant is fixed to a side of the first sealing cover facing the sealing cavity.

[0063] In some embodiments, the first sealing cover is provided with a receiving groove for receiving the sealant, and the sealant is fixed in the receiving groove. The opening of the receiving groove may face downward.

[0064] In some embodiments, the sealant is fixed to the middle or upper portion of the sealed cavity, so that the sealant can flow down to the surface of the reagent at the bottom of the sealed cavity after liquefaction to form a liquid sealing layer.

[0065] In some embodiments, the second sealing cover is configured to re-form a sealed cavity with the container body after the penetrable area of ​​the first sealing cover is penetrated.

[0066] In some embodiments, the second sealing cover is configured to re-form a sealed cavity with the container body after the penetrable area of ​​the first sealing cover is penetrated and without removing the first sealing cover.

[0067] In some embodiments, after the second sealing cover and the container body form a sealed cavity, at least a portion or the entirety of the first sealing cover is sealed in the newly formed sealed cavity.

[0068] In some embodiments, the sealing fit is an interference fit or a threaded fit.

[0069] In some embodiments, reagents for nucleic acid amplification reaction are pre-installed in the sealed cavity.

[0070] In some embodiments, the first sealing cover and the second sealing cover are two different sealing covers.

[0071] In some embodiments, the first sealing cover and / or the second sealing cover are made of a polymer material. Optionally, the first sealing cover and / or the second sealing cover are made of an elastic polymer material. The elastic modulus of the elastic polymer material may be less than or equal to 0.1×10 5 MPa, for example, less than or equal to 0.05×10 5 MPa.

[0072] In some embodiments, the container body can be generally cylindrical tube / bottle-shaped with a substantially circular opening at one end.

[0073] In some embodiments, a cross-section (a cross-section perpendicular to the axis) of the first sealing cover and / or the second sealing cover may be substantially circular in shape.

[0074] In some embodiments, the sealing fit is a circumferentially sealing fit.

[0075] In some embodiments, the sealing fit between the first sealing cover and the side wall includes: circumferential sealing fit between the outer edge of the first sealing cover and the inner edge of the side wall and / or circumferential sealing fit between the inner edge of the first sealing cover and the outer edge of the side wall.

[0076] In some embodiments, the sealing fit between the second sealing cover and the side wall includes: circumferential sealing fit between the outer edge of the second sealing cover and the inner edge of the side wall, and / or circumferential sealing fit between the inner edge of the second sealing cover and the outer edge of the side wall.

[0077] In some embodiments, the sealing fit between the second sealing cover and the first sealing cover includes: circumferential sealing fit between the outer edge of the second sealing cover and the inner edge of the first sealing cover, and / or circumferential sealing fit between the inner edge of the second sealing cover and the outer edge of the first sealing cover.

[0078] In some embodiments, the reagent is a liquid reagent or a solid reagent.

[0079] In some embodiments, the reagent is a nucleic acid amplification reagent. The reagent may contain any component for nucleic acid amplification. The reagent may be selected from template nucleic acid, primer, enzyme (such as DNA polymerase, reverse transcriptase), auxiliary factor (such as monovalent or divalent cation, such as Mg 2+ ), deoxynucleoside triphosphates (dNTPs), a buffer, and a solvent.

[0080] In some aspects, a method for operating a reagent container is provided, comprising the steps of:

[0081] a) providing any one of the reagent containers described above;

[0082] b) penetrating the penetrable area on the first sealing cover to inject a reagent into the container body or remove a reagent from the container body;

[0083] c) Changing environmental conditions to liquefy the sealant and form a liquid sealing layer inside the container body.

[0084] d) resealing the container body with a second sealing cover;

[0085] Optionally, also include

[0086] f) Changing environmental conditions to solidify the liquid sealing layer to form a solid sealing layer.

[0087] Based on the above solution, the reagent can be easily injected into the container body or taken out from the container body, and the container can also be double-sealed.

[0088] In some aspects, a method of amplifying a nucleic acid is provided, comprising:

[0089] a) providing any one of the reagent containers described above;

[0090] b) penetrating the penetrable area on the first sealing cover and injecting reagents required for the nucleic acid amplification reaction into the container body;

[0091] c) changing the ambient temperature conditions to liquefy the sealant and form a liquid sealing layer in the container body;

[0092] d) resealing the container body with a second sealing cover;

[0093] d) amplifying the nucleic acid;

[0094] Optionally, also include

[0095] f) Changing environmental conditions to solidify the liquid sealing layer to form a solid sealing layer.

[0096] Methods for using nucleic acid amplification are known in the art. The amplification reaction can be a polymerase-mediated extension reaction, such as polymerase chain reaction (PCR). However, any amplification reaction can be applicable to the disclosed scheme.

[0097] In some aspects, a reagent processing system is provided, comprising:

[0098] -reagent containers;

[0099] - a penetration mechanism, the penetration mechanism being used to penetrate the penetrable area on the first sealing cover;

[0100] Optionally, the penetration mechanism is further provided with a reagent supply mechanism and / or a reagent extraction mechanism;

[0101] - an environmental condition control mechanism, the environmental condition control mechanism is used to change the environmental conditions around the reagent container and thus change the solid-liquid state of the sealant.

[0102] In some embodiments, the reagent processing system further comprises a gripping mechanism for gripping and moving the second sealing cap.

[0103] In some embodiments, the grasping mechanism includes a limiting base, on which a puncture needle is disposed.

[0104] In some embodiments, the gripping mechanism includes a base and a gripping protrusion located on the base, and the second sealing cover is provided with a matching recess capable of interference fit with the protrusion;

[0105] The base is further provided with a retractable top block, which is located beside the convex portion. When the top block is extended, it can push down the second sealing cover which is in interference fit with the convex portion from the convex portion.

[0106] In some embodiments, the reagent processing system is a system for nucleic acid amplification.

[0107] In some embodiments, the environmental condition control mechanism is a thermal cycling instrument.

[0108] Terminology Notes:

[0109] If the present invention uses the following terms, they may have the following meanings:

[0110] Various relative terms, such as "front," "back," "top," and "bottom," "upper," "lower," "above," and "below," may be used to facilitate the description of various embodiments. Relative terms are defined with respect to a general orientation of a structure and do not necessarily indicate the actual orientation of the structure when manufactured or used.

[0111] As used in the description and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0112] The term "container" refers to a vessel suitable for receiving, storing, transporting and / or releasing contents such as test samples (e.g., blood, urine, serum, plasma or liquefied biopsy samples, etc.), test reagents (e.g., reagents for immunochemical tests, clinical chemistry tests, coagulation tests, hematology tests, molecular biology tests, etc.), or a combination thereof.

[0113] In one embodiment, the container can be a vessel with a cylindrical, conical or cubic shape. The container can have a closed bottom and an open top. The closed bottom of the cylindrical vessel can be circular, and the open top can be closable, for example, by using a lid. The non-limiting example of a single cylindrical or conical separation vessel is a primary or secondary container well known in the art. Alternatively, two or more containers can be arranged as a multi-container assembly. The non-limiting example of this multi-container assembly is a porous plate, which is well known in the art.

[0114] The term "nucleic acid amplification" generally refers to a technique for increasing the copy number of a nucleic acid molecule in a sample or specimen. Techniques that can be used for nucleic acid amplification are well known in the art. An example of nucleic acid amplification is polymerase chain reaction (PCR), in which a nucleic acid sample collected from a subject is contacted with a primer (the nucleic acid sample can be single-stranded or double-stranded, and if the nucleic acid sample is double-stranded, the double strand is first dissociated and then annealed and contacted with the primer) under conditions that allow hybridization of the primer to the nucleic acid template in the sample, the primer is extended under suitable conditions, and then the steps of dissociation (denaturation), annealing and extension are repeated to amplify the copy number of the nucleic acid. Other examples of in vitro amplification techniques include strand displacement amplification, non-transcriptional isothermal amplification, repair chain reaction amplification, ligase chain reaction, gap-filling ligase chain reaction amplification, coupled ligase detection and PCR, and RNA non-transcriptional amplification.

[0115] The term "nucleic acid" generally refers to a polymeric form of nucleotides (deoxyribonucleotides (dNTPs) or ribonucleotides (rNTPs)) of any length or their analogs. The nucleic acid may have any three-dimensional structure and may perform any known or unknown function. For the inventions involved in this application, non-limiting examples of nucleic acids include DNA, RNA, coding or non-coding regions of a gene or gene fragment, one or more loci determined by linkage analysis, exons, introns, messenger RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), short interfering RNA (siRNA), short hairpin RNA (shRNA), microRNA (miRNA), ribozymes, cDNA, recombinant nucleic acids, branched nucleic acids, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. Nucleic acids may contain one or more modified nucleotides, such as methylated nucleotides and nucleotide analogs. The nucleotide sequence of a nucleic acid may be interrupted by non-nucleotide components. Nucleic acids may be further modified after polymerization (e.g., by coupling or binding to a reporter).

[0116] The term "thermal cycling" generally refers to the process of repeatedly changing a reaction system (such as a mixture of chemical reactions) between two or more different temperatures.

[0117] The term "wax" is used broadly to encompass any kind of conventional or non-conventional wax and any artificial or natural wax, as well as any other material (whether or not called a wax) that undergoes a reversible phase transition from solid to liquid at a temperature in the range of, for example, 20°C to 100°C. When the term "wax" is used, it includes a single wax or any mixture of waxes in any proportion.

[0118] Beneficial effects

[0119] One or more technical solutions disclosed herein have one or more of the following beneficial effects:

[0120] (1) When adding / removing reagents, there is no need to open the container cover, which reduces the difficulty of operation, improves the operating efficiency, and facilitates automated operation;

[0121] (2) During the use of the container, in addition to the seal between the container cover and the container wall, additional sealing measures are provided to reduce the risk of reagent leakage;

[0122] (3) After the container is used, in addition to the seal between the container lid and the container wall, additional sealing measures are provided to reduce the risk of reagent leakage. BRIEF DESCRIPTION OF THE DRAWINGS

[0123] Figure 1 is a schematic diagram of a reagent container in some embodiments;

[0124] Figure 2A schematic diagram showing a penetrable portion of a first sealing cover being penetrated by a pipette;

[0125] Figure 3 A schematic diagram showing a sealant liquefying to form a liquid sealing layer;

[0126] Figure 4 A schematic diagram showing a reagent handling system;

[0127] Figure 5 A schematic diagram showing yet another reagent processing system;

[0128] Figure 6 A schematic diagram showing yet another reagent processing system;

[0129] Figure 7 Schematic diagrams of reagent containers according to some further embodiments. DETAILED DESCRIPTION

[0130] The embodiments of the present invention will be described in detail below with reference to the examples, but it will be understood by those skilled in the art that the following examples are merely illustrative of the present invention and should not be construed as limiting the scope of the invention. Where specific conditions are not specified in the examples, the methods were performed according to conventional conditions or the conditions recommended by the manufacturer. Where the manufacturers of the reagents or instruments are not specified, they are all conventional products that can be obtained commercially.

[0131] Figure 1 Schematic diagram of a reagent container according to some embodiments.

[0132] In some embodiments, as Figure 1 As shown, the reagent container 1 includes a container body 10, a first sealing cover 20, and a second sealing cover 40. The container body 10 includes a bottom 11 and a sidewall 12, which define a cavity. The first sealing cover 20 is capable of sealingly cooperating with the sidewall 12, thereby defining a sealed cavity 13 together with the sidewall 12 and the bottom 11. The first sealing cover 20 is provided with a mating recess 24 for sealingly cooperating with the second sealing cover. The mating recess 24 opens upward, and the bottom of the mating recess 24 has a penetrable area 23.

[0133] like Figure 1 As shown, the second sealing cover 40 is in a separated state. The second sealing cover 40 is completely separated from the container body 10 and does not seal the container body 10. The second sealing cover can also be in a mating state. When the second sealing cover 40 is in the mating state ( Figure 1 The second sealing cover 40 is interference-fitted with the matching recess 24 , thereby forming a sealed cavity together with the first sealing cover and the container body 10 .

[0134] like Figure 1As shown, the reagent container further includes a sealant 30. The sealant 30 is located in the sealed cavity 13 and is fixed to the first sealing cover 20. Specifically, the first sealing cover 20 is provided with a receiving groove 21 for receiving the sealant 30, and the sealant 30 is fixed in the receiving groove 21. The opening 22 of the receiving groove 21 faces downward.

[0135] Figure 2 Schematic diagram showing the penetrable portion of a first sealing cap being penetrated by a pipette.

[0136] like Figure 2 As shown, the pipette 50 penetrates the penetrable portion 23 of the first sealing cover 20 and extends into the first sealed cavity 13. At this time, the pipette 50 can inject reagents into the first sealed cavity 13 or take reagents out of the first sealed cavity 13.

[0137] Figure 3 Schematic diagram showing the liquefaction of a sealant to form a liquid sealing layer.

[0138] like Figure 3 As shown, the solid sealant 30 liquefies due to changes in environmental conditions, and then flows down from the opening 22 of the receiving groove 21 to the lower part of the container body 10, forming a liquid sealing layer 31 on the surface of the reagent.

[0139] During the nucleic acid amplification process, the sealant 30 can be liquefied and flowed down at an appropriate time as needed to form a liquid sealing layer. The liquid sealing layer can well seal the reaction system and prevent the reagents and the aerosols formed by the reagents from overflowing the container during the reaction.

[0140] Figure 4 A schematic diagram showing a system for nucleic acid amplification.

[0141] like Figure 4 As shown, the nucleic acid amplification system includes a reagent container 1, a penetration mechanism 50, and an environmental condition control mechanism 70. As described above, the first sealing cap 20 of the reagent container 1 is provided with a penetrable region 23. The penetration mechanism 50 is configured to penetrate the penetrable region 23. The penetration mechanism 50 includes a reagent supply mechanism that can inject the reagent into the container body 10 after penetrating the penetrable region 23. The environmental condition control mechanism 70 is configured to change the environmental conditions near the reagent container 1, thereby altering the solid / liquid state of the sealant 30.

[0142] Figure 4 (1) to (3) show the process of injecting reagent once and forming a liquid sealing layer. Figure 4As shown in (1), the reagent container 1 includes a container body 10 and a first sealing cover 20. The first sealing cover 20 is sealed with the side wall of the container body 10 to form a sealed cavity 13. A penetrable area 23 is provided on the first sealing cover. A sealant 30 is provided on the upper part of the sealed cavity 13, and a reagent 80 is pre-placed at the bottom of the sealed cavity 13. Figure 4 As shown in (2), the penetration mechanism 50 penetrates the penetrable area 23 and injects the reagent into the container body 10. Figure 4 As shown in (3), the environmental condition control mechanism 70 is located next to the reagent container 1. The environmental condition control mechanism 70 changes the environmental conditions to liquefy the sealant 30 and leave it, thereby forming a liquid sealing layer 31. In one embodiment, the environmental condition control mechanism 70 is a temperature regulating device, such as a heating device / cooling device; changing the environmental conditions can be changing the temperature, such as increasing the temperature / lowering the temperature.

[0143] Figure 5 A schematic diagram showing yet another nucleic acid amplification system.

[0144] like Figure 5 As shown, the nucleic acid amplification system further includes a grabbing mechanism 60. The grabbing mechanism 60 includes a limiting base 61, on which a puncture needle 62 is provided. The puncture needle 62 can penetrate into the second sealing cover 40, thereby lifting and transferring the second sealing cover 40.

[0145] Figure 5 (1) to (3) show a process of using the gripping mechanism 60 to grip the sealing cap 40 and seal the container body 10 therewith.

[0146] like Figure 5 As shown, in step (1), the puncture needle 62 can penetrate the second sealing cover 40, so that the second sealing cover 40 can be lifted and transferred. In step (2), the puncture needle 62 carries the second sealing cover 40 to the top of the container body 10, and inserts the second sealing cover 40 into the matching recess 24 of the first sealing cover 20 so that the two are interference fit. The limiting base 61 can prevent the second sealing cover from continuing to move upward along the puncture needle. After the first sealing cover 20 and the second sealing cover 40 are interference fit, the puncture needle 62 is pulled out. At this time, the second sealing cover seals the cavity of the container body again.

[0147] Figure 6 A schematic diagram showing yet another portion of a reagent handling system.

[0148] like Figure 6As shown, the nucleic acid amplification system further includes a gripping mechanism 60. The gripping mechanism 60 includes a base 61 and a gripping protrusion 64 located on the base. The second sealing cover 40 is provided with a gripping recess 41 that is capable of an interference fit with the gripping protrusion 64. The base 61 is also provided with a retractable push block 63, located adjacent to the protrusion 64. When extended, the push block 63 pushes down the second sealing cover 40, which is in an interference fit with the gripping protrusion 64, thereby disengaging the gripping protrusion.

[0149] Figure 6 (1) to (4) show a process of using the gripping mechanism 60 to grip the second sealing cap 40 and seal the container body 10 with it.

[0150] like Figure 6 As shown, in step (1), the gripping protrusion 64 of the gripping mechanism 60 is interference-fitted with the gripping recess 41 of the second sealing cover 40 to grip and transfer the second sealing cover 40. In step (2), the gripping mechanism 60 carries the second sealing cover 40 to the top of the first sealing cover 40 of the container body 10, and inserts the second sealing cover 40 into the matching recess 24 of the first sealing cover 20 (the matching recess 24 is as shown in FIG. Figure 3 After the first sealing cover 20 and the second sealing cover 40 have achieved an interference fit, the push block 63 is extended to push the second sealing cover 40 off the gripping protrusion 64. At this point, the second sealing cover 40 reseals the cavity of the container body 10.

[0151] Figure 7 Schematic diagrams of reagent containers according to some further embodiments.

[0152] like Figure 7 As shown, the reagent container includes a container body 10, a first sealing cover 720, and a second sealing cover 740. The container body 10 includes a bottom 11 and sidewalls 12, which define a cavity. The first sealing cover 20 seals against the sidewalls 12, thereby defining a sealed cavity 13 together with the sidewalls 12 and the bottom 11. The first sealing cover 20 is located below the upper edge of the sidewalls. The top of the first sealing cover 720 is lower than the upper edge of the sidewalls, leaving a reserved space from the upper edge of the sidewalls for accommodating the second sealing cover. The first sealing cover 20 is provided with a permeable area 23. The second sealing cover 40 can be in a detached state and a mated state. When in the detached state, the second sealing cover 40 is separated from the container body 10. When in the mated state, the outer edge of the second sealing cover 40 seals against the sidewalls of the container body 10. The reagent container also includes a sealant 730, which is located within the sealed cavity 13 and adheres to the sidewalls 12. The sealant 730 can undergo a solid-liquid phase transition as environmental conditions change.

[0153] Although the specific embodiments of the present invention have been described in detail, those skilled in the art will understand that various modifications and changes can be made to the details based on all the teachings disclosed, and these changes are all within the scope of protection of the present invention. The full scope of the present invention is given by the appended claims and any equivalents thereof.

Claims

1. A reagent processing system comprising: - a reagent container, the reagent container being a container for nucleic acid amplification, comprising: (a) a container body, the container body comprising a bottom and side walls, the bottom and side walls defining a cavity; (b) a first sealing cover, the first sealing cover being capable of sealingly cooperating with the side wall, thereby defining a sealed cavity together with the side wall and the bottom of the container body; (c) a penetrable area is provided on the first sealing cover, wherein the penetrable area is a pierceable membrane; (d) sealant wax, and The sealant wax is positioned so that when the first sealing cover, the side wall, and the bottom jointly define a sealed cavity, the sealant wax is located within the sealed cavity, and the sealant wax is fixed to the side wall or to a side of the first sealing cover facing the sealed cavity; The sealant wax can undergo a solid-liquid phase transition as the temperature changes. The sealant wax is solid at temperature T1 and liquid at temperature T2, where T2>T1; (e) a second sealing cover, wherein the second sealing cover can be in a separated state and a mated state; the separated state means that the second sealing cover does not form a sealed cavity with the container body; the mated state means that the second sealing cover and the container body define a sealed cavity; - a penetration mechanism, the penetration mechanism being used to penetrate the penetrable area on the first sealing cover; - An environmental condition control mechanism, which is used to change the environmental conditions around the reagent container and thereby change the solid-liquid state of the sealant, including: changing the environmental temperature condition to liquefy the sealant and form a liquid sealing layer within the container body; and also changing the environmental temperature condition to solidify the liquid sealing layer and form a solid sealing layer.

2. The reagent processing system according to claim 1, wherein The reagent container is a PCR tube.

3. The reagent processing system according to claim 1, wherein: The mating state means that the second sealing cover is sealed and mated with the side wall or the first sealing cover, thereby defining a sealed cavity with the container body.

4. The reagent processing system according to claim 1, wherein: The penetration mechanism is also provided with a reagent supply mechanism and / or a reagent extraction mechanism.

5. The reagent processing system according to claim 1, wherein: The environmental condition control mechanism is a thermal cycler.

6. The reagent processing system according to claim 1, characterized in that Any of the following: - when the first sealing cover, the side wall and the bottom of the container body jointly define a sealed cavity, the height of the top of the first sealing cover is lower than the height of the upper edge of the side wall; or - The first sealing cover is provided with a matching portion capable of sealingly matching with the second sealing cover.

7. The reagent processing system according to claim 6, characterized in that Any of the following: - a matching recess is provided on the first sealing cover, and the penetrable area is provided at the bottom of the matching recess of the first sealing cover; or The first sealing cover is provided with a matching protrusion, the second sealing cover is correspondingly provided with a matching recess, and the penetrable area is provided on the top of the matching protrusion of the first sealing cover. 8 . The reagent processing system according to claim 1 , wherein the second sealing cover is configured to re-form a sealed cavity with the container body after the penetrable area of ​​the first sealing cover is penetrated.

9. The reagent processing system according to claim 1, wherein the sealing fit is an interference fit or a threaded fit. 10 . The reagent processing system according to claim 1 , wherein when the second sealing cover is in the separated state, the second sealing cover is completely separated from the container body. The reagent processing system according to claim 1 , wherein reagents for nucleic acid amplification reaction are pre-installed in the sealed chamber. 12 . The reagent processing system according to claim 1 , further comprising a gripping mechanism for gripping and moving the second sealing cover.

13. The reagent processing system according to claim 12, wherein: The grabbing mechanism comprises a limiting base, and a puncture needle is arranged on the limiting base.

14. The reagent processing system according to claim 12, wherein: The grabbing mechanism includes a base, the base is provided with a grabbing protrusion, and the second sealing cover is provided with a grabbing recess capable of interference fit with the grabbing protrusion; The base is also provided with a retractable top block, which is located beside the grabbing protrusion. When the top block is extended, the second sealing cover which is in interference fit with the grabbing protrusion can be pushed down from the grabbing protrusion.

15. A method for operating a reagent container, comprising the following steps: a) providing a reagent container for nucleic acid amplification, the reagent container comprising: a container body, the container body comprising a bottom and side walls, the bottom and side walls defining a cavity; a first sealing cover, the first sealing cover being capable of sealingly cooperating with the side wall, thereby defining a sealed cavity together with the side wall and the bottom of the container body; The first sealing cover is provided with a penetrable area, wherein the penetrable area is a pierceable membrane; --sealant wax, and The sealant wax is positioned so that when the first sealing cover, the side wall, and the bottom together define a sealed cavity, the sealant wax is located within the sealed cavity; and the sealant wax is fixed to the side wall or to a side of the first sealing cover facing the sealed cavity; The sealant wax can undergo a solid-liquid phase transition as the temperature changes. The sealant wax is solid at temperature T1 and liquid at temperature T2, where T2>T1; a second sealing cover, the second sealing cover being capable of being in a detached state and a mated state; the detached state means that the second sealing cover does not form a sealed cavity with the container body; the mated state means that the second sealing cover and the container body define a sealed cavity; b) penetrating the penetrable area on the first sealing cover to inject a reagent into the container body or remove a reagent from the container body; c) changing the ambient temperature to liquefy the sealant and form a liquid sealing layer in the container body; d) resealing the container body with a second sealing cover; Also includes f) Changing the ambient temperature to solidify the liquid sealing layer and form a solid sealing layer.

16. The method according to claim 15, wherein The nucleic acid amplification reagent container is a PCR tube.

17. The method according to claim 15, wherein: The mating state means that the second sealing cover is sealed and mated with the side wall or the first sealing cover, thereby defining a sealed cavity with the container body.

18. The method according to any one of claims 15 to 17, characterized in that Any of the following: - when the first sealing cover, the side wall and the bottom of the container body jointly define a sealed cavity, the height of the top of the first sealing cover is lower than the height of the upper edge of the side wall; or - The first sealing cover is provided with a matching portion capable of sealingly matching with the second sealing cover.

19. The method according to claim 18, characterized in that Any of the following: - a matching recess is provided on the first sealing cover, and the penetrable area is provided at the bottom of the matching recess of the first sealing cover; or The first sealing cover is provided with a matching protrusion, the second sealing cover is correspondingly provided with a matching recess, and the penetrable area is provided on the top of the matching protrusion of the first sealing cover.

20. The method according to any one of claims 15 to 17, wherein the second sealing cover is configured to re-form a sealed cavity with the container body after the penetrable area of ​​the first sealing cover is penetrated.

21. The method according to any one of claims 15 to 17, wherein the sealing fit is an interference fit or a threaded fit.

22. The method according to any one of claims 15 to 17, wherein when the second sealing cover is in the separated state, the second sealing cover is completely separated from the container body.

23. The method according to any one of claims 15 to 17, wherein reagents for nucleic acid amplification reaction are pre-installed in the sealed cavity.

24. A method for amplifying nucleic acid, comprising: a) providing a nucleic acid amplification reagent container, the reagent container comprising: a container body, the container body comprising a bottom and side walls, the bottom and side walls defining a cavity; a first sealing cover, the first sealing cover being capable of sealingly cooperating with the side wall, thereby defining a sealed cavity together with the side wall and the bottom of the container body; The first sealing cover is provided with a penetrable area, wherein the penetrable area is a pierceable membrane; --sealant wax, and The sealant wax is positioned so that when the first sealing cover, the side wall, and the bottom together define a sealed cavity, the sealant wax is located within the sealed cavity; and the sealant wax is fixed to the side wall or to a side of the first sealing cover facing the sealed cavity; The sealant wax can undergo a solid-liquid phase transition as the temperature changes. The sealant wax is solid at temperature T1 and liquid at temperature T2, where T2>T1; a second sealing cover, the second sealing cover being capable of being in a detached state and a mated state; the detached state means that the second sealing cover does not form a sealed cavity with the container body; the mated state means that the second sealing cover and the container body define a sealed cavity; b) penetrating the penetrable area on the first sealing cover and injecting reagents required for the nucleic acid amplification reaction into the container body; c) changing the ambient temperature conditions to liquefy the sealant and form a liquid sealing layer in the container body; d) resealing the container body with a second sealing cover; d) amplifying the nucleic acid; Also includes f) Changing the ambient temperature to solidify the liquid sealing layer and form a solid sealing layer.

25. The method according to claim 24, wherein The reagent container is a PCR tube.

26. The method according to claim 24, wherein The mating state means that the second sealing cover is sealed and mated with the side wall or the first sealing cover, thereby defining a sealed cavity with the container body.

27. The method according to any one of claims 24 to 26, characterized in that Any of the following: - when the first sealing cover, the side wall and the bottom of the container body jointly define a sealed cavity, the height of the top of the first sealing cover is lower than the height of the upper edge of the side wall; or - The first sealing cover is provided with a matching portion capable of sealingly matching with the second sealing cover.

28. The method according to claim 27, characterized in that Any of the following: - a matching recess is provided on the first sealing cover, and the penetrable area is provided at the bottom of the matching recess; or The first sealing cover is provided with a matching protrusion, and the second sealing cover is correspondingly provided with a matching recess, and the penetrable area is provided on the top of the matching protrusion of the first sealing cover.

29. The method according to any one of claims 24 to 26, wherein the second sealing cover is configured to re-form a sealed cavity with the container body after the penetrable area of ​​the first sealing cover is penetrated.

30. The method according to any one of claims 24 to 26, wherein the sealing fit is an interference fit or a threaded fit.

31. The method according to any one of claims 24 to 26, wherein when the second sealing cover is in the separated state, the second sealing cover is completely separated from the container body.

32. The method according to any one of claims 24 to 26, wherein reagents for nucleic acid amplification reaction are pre-installed in the sealed cavity.

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