CN113493736A - Reagent container and method for operating the container - Google Patents

Abstract

The present invention relates to a reagent vessel and a method of operating the same, the reagent vessel comprising a vessel body, a first sealing lid, a sealant and a second sealing lid. The reagent container has improved sealing properties.

Description

Reagent container and method for operating the container

Technical Field

The present disclosure relates to the field of biological detection technology, and more particularly, to a reagent container and a method for operating the same.

Background

Polymerase Chain Reaction (PCR) is a molecular biology technique that can be used to amplify specific DNA fragments. The method can be used for carrying out in-vitro efficient amplification on the selective fragment of the gene, thereby realizing the detection of the target gene.

Because PCR products are exponentially accumulated in the amplification process, after the PCR reaction is finished, the concentration of the products in the PCR tube is tens of millions of times or even higher than that before the reaction, and once the PCR tube is not tightly sealed, the high-concentration products form aerosol and leak, so that serious pollution is caused, and subsequent PCR detection is abnormal.

In the operation of nucleic acid amplification, it is sometimes necessary to add or remove a part of the preparation to or from the container, and it is necessary to open the lid of the reagent container and close the lid after adding or removing the reagent.

Disclosure of Invention

The inventors have found that the following problems exist with the existing reagent containers:

(1) the container is sealed only by forming mechanical seal between the container cover and the container wall, and is single seal, so that the risk of untight seal exists;

(2) since high-concentration and volatile aerosol is generated during and after the nucleic acid amplification reaction, the gas can cause laboratory pollution if leaked into a laboratory;

(3) when the reagent needs to be added/taken out, the container cover needs to be opened, and the existing scheme has difficulty in realizing automation operation. (ii) a

(4) Containers that are left ready for use after use still risk reagent leakage.

The present disclosure provides a reagent container having at least the following advantages:

(1) when the reagent needs to be added or taken out, the container cover does not need to be opened, so that the operation difficulty is reduced, the operation efficiency is improved, and the automatic operation is facilitated;

(2) in the use process of the container, besides the sealing between the container cover and the container wall, a plurality of sealing measures are provided, so that the risk of reagent leakage in the container is reduced;

(3) after the container is used and stands for use, besides the sealing between the container cover and the container wall, the container also provides one more sealing measure, and reduces the risk of reagent leakage in the container.

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

a main body of the container is provided with a plurality of grooves,

the container body includes a bottom and a sidewall defining a cavity;

a first sealing cover is arranged at the bottom of the container,

the first sealing cover can be in sealing fit with the side wall, so that a sealing cavity is defined by the first sealing cover and the side wall and the bottom of the container body;

a penetrable region is arranged on the first sealing cover;

a sealing agent is added to the mixture of the components,

the sealant is positioned such that it is within the sealed cavity when the first seal cover, sidewall, and bottom collectively define the sealed cavity;

the sealant is capable of undergoing a solid-liquid phase change as environmental conditions change; and

a second sealing cover is arranged on the first sealing cover,

the second sealing cover can be in a separated state and a matched state;

the separated state means that the second sealing cover does not form a sealing cavity with the container body;

the matched state means that the second sealing cover and the container main body define a sealing cavity.

The reagent container has the advantages that:

(1) by operating a penetration mechanism (such as a pipette), the penetrable region on the first sealing cover can be penetrated, so that the reagent can be added into or taken out from the sealing cavity without additional operation of opening the cover; therefore, the operation efficiency is improved, and the automatic operation is facilitated;

(2) by changing the environmental conditions, the sealant is liquefied to form a liquid sealing layer, and then the liquid sealing layer and the second sealing cover are combined to form a seal with the container body, so that a double-sealing system is constructed by the liquid sealing layer and the second sealing cover, and the risk of reagent leakage is greatly reduced;

(3) after the container is used, the sealant for forming the liquid sealing layer can be cured to form a solid sealing layer, and the solid sealing layer and the second sealing cover are in a double sealing system, so that the leakage risk of the reagent is greatly reduced.

In some embodiments, the engaged state is when the second sealing cap is in sealing engagement with the sidewall or the first sealing cap, thereby defining a sealed cavity with the container body.

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 chamber is broken, which communicates with the external environment, thereby enabling the step of adding/removing reagents.

In some embodiments, the penetrable region may be located in a central region of the first sealing cap at a location that facilitates penetration. The area of the penetrable region can account for 1-100% of the cross-sectional area of the first sealing cover.

In some embodiments, the penetrable region of the first sealing cap is a pierceable membrane.

In some embodiments, the penetrable region of the first sealing cap is a plastic film or a metal foil.

In some embodiments, the second sealing cap is completely separated from the container body when the second sealing cap is in the separated state.

In some embodiments, the container body may be resealed by adjusting the second seal cap to a mated condition after the penetrable region of the first seal cap is penetrated.

In some embodiments, the second sealing cap sealingly engages the sidewall of the container body when the second sealing cap is in engagement, with the second sealing cap and the container body defining a second sealed cavity. At this time, the penetrated first sealing cap may be sealed in the second sealing chamber.

In some embodiments, the second seal cap is in sealing engagement with the first container cap when the second seal cap is in engagement, with the second seal cap, the first seal cap and the container body defining a second seal cavity. At this point, the penetrated penetrable region may be sealed within the second sealed chamber.

In some embodiments, when the first sealing cover and the sidewall and bottom of the container body together 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 sidewall. Based on this, the top of the first sealing cover is reserved to the upper edge of the side wall. After the penetrable region of the first sealing cover is penetrated, a sealing cavity can be formed with the container body again by covering the reserved space with the second sealing cover.

In some embodiments, the first sealing cover is provided with a matching portion (e.g., a matching surface, a matching concave portion, a matching convex portion or an annular matching portion, such as a groove, an annular wall, an annular step surface, etc.) capable of being in sealing matching with the second sealing cover, and correspondingly, the second sealing cover can also be provided with a matching portion (e.g., a matching surface, a matching concave portion, a matching convex portion or an annular matching portion) capable of being in sealing matching with the first sealing cover. Therefore, after the penetrable region on the first sealing cover is penetrated, the penetrated penetrable region can be blocked by the sealing and matching of the second sealing cover and the first sealing cover, and the resealing of the sealing cavity is realized.

In some embodiments, a mating recess is provided on the first sealing cap. The opening of the mating recess may be upward, the mating recess may include a mating recess sidewall and a bottom, and the penetrable region is disposed at the bottom of the mating recess. At the moment, the outer edge of the second sealing cover is in circumferential sealing fit with the inner side of the side wall of the matching concave part, so that the penetrated penetrable region can be sealed and blocked, and the sealing cavity is sealed again.

In some embodiments, a mating protrusion is provided on the first sealing cap and a mating recess is correspondingly provided on the second sealing cap. The protruding direction of the fitting projection may be upward. The mating projection may include a mating projection sidewall and a top, with the penetrable region disposed at the top of the mating projection. At the moment, the matching concave part of the second sealing cover is in circumferential sealing fit with the outer side of the side wall of the matching convex part of the first sealing cover, so that the penetrated penetrable region can be sealed and blocked, and the sealing cavity is sealed again.

In some embodiments, in a non-use state of the nucleic acid amplification vessel, the sealant is in a solid state; in the use state of the nucleic acid amplification container, the environmental conditions can be changed as needed, and the solid sealant is liquefied to form a liquid sealant (liquid sealant) in the container main body, thereby preventing the reagent and the aerogel formed by the reagent from overflowing the container during the reaction. After the reaction is completed, the liquid sealing layer may be cured to form a solid seal (solid sealing layer) by changing the environmental conditions as necessary.

In some embodiments, the formation of a liquid-tight layer on the surface of an existing formulation in a container is achieved by changing the environmental conditions (e.g., temperature, light, or chemical environment) after all of the required reagents have been added to the nucleic acid amplification reaction to change the sealant from a solid state to a liquid state and flow to the reagent surface, thereby preventing the formulation from overflowing during subsequent reactions. After the nucleic acid amplification reaction is finished, the sealant is changed from a liquid state to a solid state to form a solid seal by changing the environmental conditions, so that the leakage of the preparation inside the container after standing for use is avoided.

In some embodiments, the change in environmental conditions refers to a change in temperature, a change in lighting conditions, or a change in chemical environment.

In some embodiments, the sealant is a solid below temperature T1 (including T1) and a liquid above temperature T2 (including T2), T2 > T1. At this time, the sealant can be liquefied by heating the container body, and a liquid seal layer can be formed in the tube. By cooling the container body, the sealant of the liquid sealant layer can be solidified to form a solid closure.

In some embodiments, T1 is 20 to 40 ℃, e.g., 30 to 40 ℃.

In some embodiments, T2 is 45 to 95 ℃, e.g., 60 to 90 ℃, e.g., 70 to 80 ℃.

In some embodiments, the control of the solid-liquid state of the sealant can be achieved by applying thermal radiation and optical radiation to the sealant located in the container body from the outside of the container body through a temperature control device, an illumination device and the like.

In some embodiments, the density of the sealant is less than the density of water.

In some embodiments, the sealant is insoluble in water.

In some embodiments, the sealant is secured to the first closure cap or within the container body.

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

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

In certain embodiments, the sealing reagent is a solid and is capable of being converted to a liquid under predetermined conditions, and the liquid has a density that is less than the density of the reagent for nucleic acid amplification, and the liquid is immiscible with the reagent for nucleic acid amplification (e.g., it is immiscible with an aqueous solution and is not volatile). In certain embodiments, the predetermined condition is selected from heating, light irradiation and/or chemical treatment. For example, in certain embodiments, the sealing reagent is a solid and is capable of being converted to a liquid under heating conditions, and the liquid has a density that is less than the density of the nucleic acid amplification reagent, and the liquid is immiscible with the nucleic acid amplification reagent (e.g., it is immiscible with an aqueous solution and is not volatile).

As used herein, the term "immiscible" means that two or more liquids do not dissolve in each other when placed in the same container, and spontaneously create a boundary interface, forming a layer when they come into contact.

As used herein, the term "layering" refers to the spontaneous formation of a vertically distributed structure due to the density difference when two or more immiscible liquids are placed in the same container, and the liquid with high density sinks while the liquid with low density floats upwards to form a layer.

As used herein, the sealing reagent of the present invention is immiscible with the reagent for nucleic acid amplification in a liquid state, and has a density lower than that of the reagent for nucleic acid amplification. 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 on the upper layer, and isolate (i.e., seal) the nucleic acid amplification reagent (and other substances, such as nucleic acids, that may be contained therein) located on the lower layer, thereby preventing both the reagents of the lower layer (and other substances, such as nucleic acids, that may be contained therein) from escaping to the external environment and the substances in the external environment from entering and contaminating the lower layer. In the present application, it is preferred that the nucleic acid molecule is not soluble in the sealing reagent of the present invention. Therefore, the sealing agent of the present invention can exert a better isolation effect. Further, it is to be understood that any substance capable of achieving this function may be used as the sealing agent of the present application, and therefore, the sealing agent of the present application is not limited to the substances listed in the present disclosure.

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

In some embodiments, the sealant is affixed to the sidewall.

In some embodiments, the sealant is secured to a side of the first seal cover facing the sealed cavity.

In some embodiments, a receiving groove for receiving a sealant is provided on the first sealing cap, and the sealant is fixed in the receiving groove. The opening of the receiving groove may be downward.

In some embodiments, the sealant is fixed to the middle or upper portion of the sealed chamber so that the sealant can flow down to the surface of the reagent at the bottom of the sealed chamber after liquefaction to form a liquid-tight layer.

In some embodiments, the second sealing cap is configured to be able to re-form a sealed chamber with the container body after the penetrable region of the first sealing cap is penetrated.

In some embodiments, the second sealing cap is configured to re-form a sealed chamber with the container body after the penetrable region of the first sealing cap is penetrated and without removing the first sealing cap.

In some embodiments, after the second seal cap forms a sealed cavity with the container body, at least a portion or all of the first seal cap is sealed within the reformed sealed cavity.

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

In some embodiments, the sealed chamber is pre-filled with reagents for a nucleic acid amplification reaction.

In some embodiments, the first seal cap and the second seal cap are two different seal caps.

In some embodiments, the first seal cap and/or the second seal cap are made of a polymeric material. Alternatively,the first and/or second seal covers are made of a resilient polymeric material. The elastic modulus of the elastic polymeric material can be less than or equal to 0.1 x 10⁵MPa, e.g. less than or equal to 0.05X 10⁵MPa。

In some embodiments, the container body may be substantially cylindrical tube/bottle shaped with a substantially circular opening at one end.

In some embodiments, the cross-section (a section perpendicular to the axis) of the first seal cap and/or the second seal cap may be substantially circular in shape.

In some embodiments, a sealing fit refers to a circumferentially sealing fit.

In some embodiments, the sealing engagement of the first sealing cover with the sidewall comprises: the outer edge of the first seal cap is in circumferential sealing engagement with the inner edge of the sidewall and/or the inner edge of the first seal cap is in circumferential sealing engagement with the outer edge of the sidewall.

In some embodiments, the sealing engagement of the second sealing cover with the sidewall comprises: the outer edge of the second seal cap is in circumferential sealing engagement with the inner edge of the sidewall and/or the inner edge of the second seal cap is in circumferential sealing engagement with the outer edge of the sidewall.

In some embodiments, the sealing engagement of the second seal cap with the first seal cap comprises: the outer edge of the second seal cap is in circumferential sealing engagement with the inner edge of the first seal cap and/or the inner edge of the second seal cap is in circumferential sealing engagement with the outer edge of the first seal cap.

In some embodiments, the reagent refers to a liquid reagent or a solid reagent.

In some embodiments, the reagent is a reagent for nucleic acid amplification. The reagent may contain any component for nucleic acid amplification. The reagent may be selected from the group consisting of a template nucleic acid, a primer, an enzyme (e.g., DNA polymerase, reverse transcriptase), a cofactor (e.g., a monovalent or divalent cation, such as Mg²⁺) One or more of deoxynucleoside triphosphate (dNTP), a buffer and a solvent.

In some aspects, there is provided a method of operating a reagent container, comprising the steps of:

a) providing a reagent container according to any one of the above;

b) penetrating the penetrable region on the first sealing cover, and injecting the reagent into the container body or taking the reagent out of the container body;

c) changing the environmental conditions to liquefy the sealant and form a liquid seal layer within the container body.

d) Resealing the container body with a second sealing cap;

optionally, also comprises

f) And changing the environmental conditions to solidify the liquid sealing layer to form a solid sealing layer.

Based on the scheme, the reagent can be conveniently injected into the container main body or taken out of the container main body, and the container can be sealed in a double way.

In some aspects, there is provided a method of amplifying a nucleic acid, comprising:

a) providing a reagent container according to any one of the above;

b) penetrating the penetrable region on the first sealing cover, and injecting reagents required by nucleic acid amplification reaction into the container main body;

c) changing the ambient temperature condition to liquefy the sealant and form a liquid seal layer in the container body;

d) resealing the container body with a second sealing cap;

d) amplifying the nucleic acid;

optionally, also comprises

f) And changing the environmental conditions to solidify the liquid sealing layer to form a solid sealing layer.

Methods of using nucleic acid amplification are known in the art. The amplification reaction may be a polymerase mediated extension reaction, such as the Polymerase Chain Reaction (PCR). However, any amplification reaction may be suitable for use in the disclosed protocol.

In some aspects, there is provided a reagent processing system comprising:

-a reagent container;

-a penetration means for penetrating a penetrable region on the first sealing cap;

optionally, a reagent supply mechanism and/or a reagent extraction mechanism are/is further arranged on the penetrating mechanism;

-environmental condition control means for changing the environmental conditions around the reagent container and thereby changing the solid-liquid state of the sealant.

In some embodiments, the reagent processing system further comprises a grasping mechanism for grasping and moving the second sealing lid.

In some embodiments, the grasping mechanism includes a restraining base with a puncture needle disposed thereon.

In some embodiments, the gripping mechanism comprises a base and a gripping convex part on the base, and the second sealing cover is provided with a matching concave part capable of being in interference fit with the convex part;

the base is further provided with a telescopic ejector block, the telescopic ejector block is located beside the convex part, and when the ejector block extends out, the second sealing cover in interference fit with the convex part can be ejected from the convex part.

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

In some embodiments, the environmental condition control mechanism is a thermal cycler.

Description of terms:

if the following terms are used in the present invention, they may have the following meanings:

various relative terms such as "front," "back," "top," and "bottom," "upper," "lower," "above," "below," and the like may be used to facilitate description of various embodiments. Relative terms are defined with respect to conventional orientations of the structure and do not necessarily indicate an actual orientation of the structure at the time of manufacture or use.

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.

The term "container" relates to a vessel adapted to receive, store, transport and/or release the contents such as a test sample (e.g., blood, urine, serum, plasma or liquefied biopsy sample, etc.), a test reagent (e.g., a reagent for immunochemical testing, clinical chemistry testing, coagulation testing, hematological testing, molecular biology testing, etc.), or a combination thereof.

In one embodiment, the container may be a vessel having a cylindrical, conical, or cubic shape. The container may have a closed bottom and an open top. The closed bottom of the cylindrical vessel may be circular and the open top may be closable, for example by using a lid. Non-limiting examples of a single cylindrical or conical separation vessel are primary or secondary containers as are known in the art. Alternatively, two or more containers may be arranged as a multi-container assembly. A non-limiting example of such a multi-container assembly is a multi-well plate, which is well known in the art.

The term "nucleic acid amplification" generally refers to a technique that increases the copy number of nucleic acid molecules 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 the Polymerase Chain Reaction (PCR), in which a nucleic acid sample collected from a subject is contacted with a primer (which may be single-stranded or double-stranded, if the nucleic acid sample is double-stranded, the double strand is dissociated first and then annealed in contact with the primer) under conditions that allow the primer to hybridize to a nucleic acid template in the sample, the primer is extended under suitable conditions, and the steps of dissociating (denaturing), annealing and extending are then repeated to amplify the copy number of the nucleic acid. Other examples of in vitro amplification techniques include strand displacement amplification, transcriptionless isothermal amplification, repair strand reaction amplification, ligase chain reaction, gap-fill ligase chain reaction amplification, coupled ligase detection and PCR, and RNA transcriptionless amplification, among others.

The term "nucleic acid" generally refers to a polymeric form of nucleotides of any length (deoxyribonucleotides (dntps) or ribonucleotides (rNTP)) or analogs thereof. The nucleic acid may have any three-dimensional structure and may perform any known or unknown function. Non-limiting examples of nucleic acids for the invention to which this application relates 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), micro RNA (mirna), ribozymes, cDNA, recombinant nucleic acids, branched nucleic acids, plasmids, vectors, DNA of any isolated sequence, RNA of any isolated sequence, nucleic acid probes, and primers. Nucleic acids may comprise 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. The nucleic acid may be further modified after polymerization (e.g., by coupling or binding to a reporter).

The term "thermal cycling" generally refers to the process by which a reaction system (e.g., a mixture of chemical reactions) is repeatedly varied between two or more different temperatures.

The term "wax" broadly encompasses: any kind of conventional or unconventional wax and any artificial or natural wax and any other material (whether referred to as wax) that undergoes a reversible phase change from solid to liquid at temperatures in the range of, for example, 20 deg. -100 deg.. When the term "wax" is used, its meaning includes: single wax or wax mixed arbitrarily in any proportion.

Advantageous effects

One or more technical schemes of the present disclosure have one or more of the following beneficial effects:

(1) when the reagent needs to be added or taken out, the container cover does not need to be opened, so that the operation difficulty is reduced, the operation efficiency is improved, and the automatic operation is facilitated;

(2) in the use process of the container, in addition to the sealing between the container cover and the container wall, an additional sealing measure is provided, so that the risk of reagent leakage is reduced;

(3) after the container is used, in addition to the seal between the container cover and the container wall, an additional sealing measure is provided, and the risk of reagent leakage is reduced.

Drawings

FIG. 1 is a schematic view of a reagent container of some embodiments;

FIG. 2 is a schematic view of a penetrable portion of a first sealing cap being penetrated by a pipette;

FIG. 3 shows a schematic view of a sealant being liquefied to form a liquid sealant layer;

FIG. 4 shows a schematic of a reagent processing system;

FIG. 5 shows a schematic view of yet another reagent processing system;

FIG. 6 shows a schematic view of yet another reagent processing system;

FIG. 7 is a schematic view of a reagent container according to further embodiments.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

Fig. 1 is a schematic view of a reagent container of some embodiments.

In some embodiments, as shown in fig. 1, the reagent vessel 1 includes a vessel 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, the bottom 11 and sidewall 12 defining a cavity. The first sealing cover 20 is capable of sealingly engaging the side wall 12 to define a sealed cavity 13 with the side wall 12 and the bottom 11. The first sealing cover 20 is provided with a matching concave part 24 which is used for being matched with the second sealing cover in a sealing way, the opening of the matching concave part 24 is upward, and the bottom of the matching concave part 24 is provided with a penetrable region 23.

As shown in fig. 1, the second sealing cover 40 is in a separated state, and the second sealing cover 40 is completely separated from the container body 10 without sealing the container body 10. The second seal cap can also be in a mated condition. When the second seal cover 40 is in a fitted state (not shown in fig. 1), the second seal cover 40 is interference-fitted with the fitting recess 24 to constitute a seal chamber together with the first seal cover and the container body 10.

As shown in fig. 1, the reagent vessel further includes a sealant 30. The sealant 30 is located in the sealing chamber 13 and fixed to the first sealing cap 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 is downward.

Fig. 2 shows a schematic view of the penetrable portion of a first sealing cap being penetrated by a pipette.

As shown in fig. 2, the pipette 50 penetrates the penetrable portion 23 of the first sealing cap 20 and protrudes into the first sealed chamber 13. At this time, the pipette 50 may inject a reagent into the first sealed chamber 13 or withdraw a reagent from the first sealed chamber 13.

Figure 3 shows a schematic view of a sealant liquefied to form a liquid sealant layer.

As shown in fig. 3, the solid sealant 30 liquefies due to a change in environmental conditions, and then flows down from the opening 22 of the storage tank 21 to the lower portion of the container main body 10, thereby forming a liquid sealant layer 31 on the surface of the reagent.

In the nucleic acid amplification process, the sealing agent 30 may be liquefied and flowed down at an appropriate timing as necessary to form a liquid sealing layer. The liquid sealing layer can well seal a reaction system, and reagents and aerosol formed by the reagents in the reaction process are prevented from overflowing out of the container.

FIG. 4 shows a schematic diagram of a nucleic acid amplification system.

As shown in FIG. 4, the nucleic acid amplification system comprises a reagent vessel 1, a penetration mechanism 50, and an environmental condition control mechanism 70. As mentioned above, the first sealing lid 20 of the reagent vessel 1 is provided with a penetrable region 23. The penetration mechanism 50 for penetrating the penetrable region 23 includes a reagent supply mechanism for allowing injection of a reagent into the container body 10 after penetration of the penetrable region 23. The environmental condition control mechanism 70 is used to change the environmental condition in the vicinity of the reagent vessel 1, thereby changing the solid/liquid state of the sealing agent 30.

Fig. 4 (1) to (3) show a process of injecting a reagent at a time and forming a liquid sealing layer. As shown in fig. 4 (1), the reagent vessel 1 includes a vessel body 10 and a first sealing lid 20, and the first sealing lid 20 is in sealing engagement with a side wall of the vessel body 10 to form a sealed chamber 13. The first closure cap has a penetrable region 23 thereon. The upper part of the sealed chamber 13 is provided with the sealant 30, and the bottom of the sealed chamber 13 is preset with the reagent 80. As shown in fig. 4 (2), the penetrating mechanism 50 penetrates the penetrable region 23 to inject a reagent into the container body 10. As shown in fig. 4 (3), the environmental condition control means 70 is located near the reagent container 1, and the environmental condition control means 70 liquefies and retains the sealing agent 30 by changing the environmental condition to form the liquid sealing layer 31. In one example, the environmental condition control mechanism 70 is a temperature regulating device, such as a temperature increasing device/temperature decreasing device; changing the environmental condition may be changing the temperature, e.g. increasing/decreasing the temperature.

FIG. 5 shows a schematic diagram of still another nucleic acid amplification system.

As shown in FIG. 5, the nucleic acid amplification system further includes a grasping mechanism 60. The grabbing mechanism 60 comprises a limiting base 61, and a puncture needle 62 is arranged on the limiting base 61. The piercing needle 62 can penetrate into the second sealing cap 40 so that the second sealing cap 40 can be lifted and transferred.

Fig. 5 (1) to (3) show a process of grasping the sealing cap 40 using the grasping mechanism 60 and sealing the container body 10 therewith.

As shown in fig. 5, in step (1), the piercing needle 62 may pierce the second sealing cap 40, so that the second sealing cap 40 may be lifted and transferred. In step (2), the puncture needle 62 carries the second seal cap 40 over the container body 10, and the second seal cap 40 is inserted into the fitting recess 24 of the first seal cap 20 so as to be interference-fitted. The limit base 61 prevents the second seal cap from continuing to move upward along the puncture needle. After the first seal cap 20 is in interference fit with the second seal cap 40, the puncture needle 62 is pulled out. At this time, the second sealing cap reseals the cavity of the container body.

Figure 6 shows a schematic view of a further reagent processing system portion.

As shown in FIG. 6, the nucleic acid amplification system further includes a grasping mechanism 60. The grasping mechanism 60 includes a base 61 and a grasping projection 64 on the base. The second seal cover 40 is provided with a catching recess 41 capable of interference-fitting with the catching projection 64. The base 61 is further provided with a telescopic jacking block 63, the telescopic jacking block 63 is located beside the convex part 64, and when the jacking block 63 extends out, the second sealing cover 40 in interference fit with the grabbing convex part 64 can be jacked down so as to be separated from the grabbing convex part.

Fig. 6 (1) to (4) show a flow of grasping the second seal cap 40 using the grasping mechanism 60 and sealing the container body 10 therewith.

As shown in fig. 6, in step (1), the gripping convex portion 64 of the gripping mechanism 60 grips and transfers the second seal cover 40 by interference fit with the gripping concave portion 41 of the second seal cover 40. In step (2), the gripping mechanism 60 carries the second seal cap 40 over the first seal cap 40 of the container body 10, and the second seal cap 40 is inserted into the mating recess 24 of the first seal cap 20 (the mating recess 24 is shown in fig. 3), so that the two are in interference fit. After the first sealing cover 20 is in interference fit with the second sealing cover 40, the ejector block 63 is extended, and the second sealing cover 40 is ejected from the grabbing convex part 64. At this time, the second sealing cap 40 re-seals the cavity of the container body 10.

FIG. 7 is a schematic view of a reagent container according to further embodiments.

As shown in fig. 7, the reagent vessel includes a vessel body 10, a first sealing cover 720 and a second sealing cover 740. The container body 10 includes a bottom 11 and a sidewall 12, the bottom 11 and sidewall 12 defining a cavity. The first sealing cover 20 is in sealing engagement with the side wall 12 so as to define a sealed cavity 13 together with the side wall 12 and the bottom 11. The first seal cover 20 is located below the upper edge of the side wall. The top of the first sealing cover 720 is lower than the top of the sidewall, and a space is reserved from the top of the sidewall for accommodating a second sealing cover. The first sealing cap 20 is provided with a penetrable region 23. The second sealing cover 40 can be in a separated state and an engaged state. When the second sealing cap 40 is in the separated state, it is separated from the container body 10. When the second sealing cover 40 is in the engaged state, the outer edge of the second sealing cover 40 is in sealing engagement with the sidewall of the container body 10. The reagent container further comprises a sealant 730, the sealant 730 being located within the sealed cavity 13 and attached to the sidewall 12. The encapsulant 730 is capable of a solid-liquid phase change as environmental conditions change.

While specific embodiments of the invention have been described in detail, those skilled in the art will understand that: various modifications may be made in the details within the teachings of the disclosure, and these variations are within the scope of the invention. The full scope of the invention is given by the appended claims and any equivalents thereof.

Claims (20)

1. A reagent container comprising:

a main body of the container is provided with a plurality of grooves,

the container body includes a bottom and a sidewall defining a cavity;

a first sealing cover is arranged at the bottom of the container,

the first sealing cover can be in sealing fit with the side wall, so that a sealing cavity is defined by the first sealing cover and the side wall and the bottom of the container body;

a penetrable region is arranged on the first sealing cover;

a sealant, and

the sealant is positioned such that when the first seal cover, sidewall, and bottom collectively define a sealed cavity, the sealant is within the sealed cavity;

the sealant is capable of undergoing a solid-liquid phase change as environmental conditions change;

a second sealing cover is arranged on the first sealing cover,

the second sealing cover can be in a separated state and a matched state;

the separated state means that the second sealing cover does not form a sealing cavity with the container body;

the matched state means that the second sealing cover and the container main body define a sealing cavity;

preferably, the engaged state means that the second sealing cover is in sealing engagement with the side wall or the first sealing cover so as to define a sealed cavity with the container body.

2. Reagent container according to claim 1, characterized by any of the following:

-the sealant is a wax;

-the sealant is fixed to the side wall;

-the sealant is fixed to a side of the first sealing cover facing the sealed cavity.

3. The reagent container of claim 1, wherein the change in environmental condition is a change in temperature, a change in lighting condition, or a change in chemical environment.

4. The reagent container of claim 1, the sealant being capable of a solid-liquid phase change with temperature change, the sealant being solid at a temperature T1 and liquid at a temperature T2, T2 > T1.

5. Reagent container according to claim 1, characterized by any of the following:

-when the first sealing cover defines a sealed cavity together with the side wall and the bottom of the container body, the height of the top of the first sealing cover is lower than the height of the upper edge of the side wall;

the first sealing cover is provided with a matching part which can be matched with the second sealing cover in a sealing way.

6. Reagent container according to claim 5, characterized by any of the following:

-the penetrable region is provided at the bottom of the mating recess;

-the penetrable region is provided on top of the mating protrusion.

7. A reagent container according to claim 1, the penetrable region being a pierceable membrane.

8. A reagent container according to claim 1, the second sealing cap being arranged to be able to re-form a sealed chamber with the container body after the penetrable region of the first sealing cap has been penetrated.

9. The reagent container of claim 1, the sealing fit being an interference fit or a threaded fit.

10. The reagent vessel of claim 1, wherein the second sealing cap is completely separated from the vessel body when the second sealing cap is in a separated state.

11. The reagent vessel according to claim 1, wherein a reagent for nucleic acid amplification reaction is placed in the sealed chamber.

12. The reagent vessel according to claim 1, which is a reagent vessel for nucleic acid amplification, such as a PCR tube.

13. A method of operating a reagent vessel according to any of claims 1 to 12, comprising the steps of:

a) providing a reagent container according to any one of claims 1 to 12;

b) penetrating the penetrable region on the first sealing cover, and injecting the reagent into the container body or taking the reagent out of the container body;

c) changing environmental conditions to liquefy the sealant and form a liquid seal layer in the container body;

d) resealing the container body with a second sealing cap;

optionally, also comprises

f) And changing the environmental conditions to solidify the liquid sealing layer to form a solid sealing layer.

14. A method of amplifying a nucleic acid comprising:

a) providing a reagent container according to any one of claims 1 to 12;

b) penetrating the penetrable region on the first sealing cover, and injecting reagents required by nucleic acid amplification reaction into the container main body;

c) changing the ambient temperature condition to liquefy the sealant and form a liquid seal layer in the container body;

d) resealing the container body with a second sealing cap;

d) amplifying the nucleic acid;

optionally, also comprises

f) And changing the environmental conditions to solidify the liquid sealing layer to form a solid sealing layer.

15. A reagent processing system comprising:

-a reagent container according to any one of claims 1 to 12;

-a penetration means for penetrating a penetrable region on the first sealing cap;

optionally, a reagent supply mechanism and/or a reagent extraction mechanism are/is further arranged on the penetrating mechanism;

-environmental condition control means for changing the environmental conditions around the reagent container and thereby changing the solid-liquid state of the sealant.

16. The reagent processing system of claim 15 further comprising

And the grabbing mechanism is used for grabbing and moving the second sealing cover.

17. The reagent processing system of claim 16,

the grabbing mechanism comprises a limiting base, and a puncture needle is arranged on the limiting base.

18. The reagent processing system of claim 16,

the grabbing mechanism comprises a base, a grabbing convex part is arranged on the base, and a grabbing concave part which can be in interference fit with the grabbing convex part is arranged on the second sealing cover;

the base is further provided with a telescopic ejector block, the telescopic ejector block is located beside the convex part, and when the ejector block extends out, the second sealing cover in interference fit with the convex part can be ejected from the convex part.

19. The reagent processing system of claim 15, which is a system for nucleic acid amplification.

20. The reagent processing system of claim 15 wherein the environmental condition control mechanism is a thermal cycler.

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