CN115044462B - Sampling tube capable of realizing nucleic acid amplification and detection without uncapping and application method - Google Patents

Sampling tube capable of realizing nucleic acid amplification and detection without uncapping and application method

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Publication number
CN115044462B
CN115044462B CN202210562277.8A CN202210562277A CN115044462B CN 115044462 B CN115044462 B CN 115044462B CN 202210562277 A CN202210562277 A CN 202210562277A CN 115044462 B CN115044462 B CN 115044462B
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ring
tube
tube body
cover
sampling tube
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CN115044462A (en
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张力
王焕英
武雅文
高雯雯
刘洋
刘慧君
杨斌
王庭璋
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Zhejiang Tianke High And New Technology Development Co ltd
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Zhejiang Tianke High And New Technology Development Co ltd
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/044—Connecting closures to device or container pierceable, e.g. films, membranes

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  • Bioinformatics & Cheminformatics (AREA)
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  • Clinical Laboratory Science (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

本发明提供了一种不开盖实现核酸扩增和检测的采样管及应用方法。采样管，包括管体、管盖；管体的外壁上侧从上到下设有第一突环和第二突环；管盖的内壁下方从上到下设有第一凹环和第二凹环；第一突环和第一凹环、第二凹环相适配，用于密封管体、管盖；第二突环和第二凹环相适配，用于密封管体、管盖；管盖内部从上到下分别设有废液池、分隔储液结构、插针结构；插针结构用于刺破分隔储液结构；废液池和管体之间设有排液通道，倒置时，液体从管体排到废液池，正置时，液体留存在废液池。本发明可以在不开盖情况下，通过分步下压管盖实现管盖中的反应试剂分步释放到反应体系中，实现不开盖核酸扩增和检测，避免样品间气溶胶污染和检测人员污染风险。

The present invention provides a sampling tube and application method for realizing nucleic acid amplification and detection without opening the cover. The sampling tube includes a tube body and a tube cover; a first protruding ring and a second protruding ring are provided on the upper side of the outer wall of the tube body from top to bottom; a first concave ring and a second concave ring are provided on the lower inner wall of the tube cover from top to bottom; the first protruding ring is adapted to the first concave ring and the second concave ring, and is used to seal the tube body and the tube cover; the second protruding ring and the second concave ring are adapted to seal the tube body and the tube cover; a waste liquid pool, a partition liquid storage structure, and a pin structure are respectively provided inside the tube cover from top to bottom; the pin structure is used to puncture the partition liquid storage structure; a liquid discharge channel is provided between the waste liquid pool and the tube body, and when inverted, the liquid is discharged from the tube body to the waste liquid pool, and when upright, the liquid remains in the waste liquid pool. The present invention can realize the step-by-step release of the reaction reagents in the tube cover into the reaction system by pressing down the tube cover step by step without opening the cover, so as to realize nucleic acid amplification and detection without opening the cover, and avoid the risk of aerosol contamination between samples and contamination of detection personnel.

Description

Sampling tube capable of realizing nucleic acid amplification and detection without uncapping and application method
Technical Field
The invention relates to the technical field of nucleic acid detection, in particular to a sampling tube for realizing nucleic acid amplification and detection without uncovering and an application method thereof.
Background
With the rapid development of molecular biology techniques, microorganism identification techniques have also been developed. Nucleic acid detection based on Polymerase Chain Reaction (PCR) is one of the important technological means for virus and bacteria detection.
PCR detection of viral and bacterial nucleic acids typically requires detection experiments by trained detection personnel in a specialized PCR laboratory that sets up different rooms with pressure differentials for reagent preparation, sample pretreatment and amplification analysis, respectively, depending on the experimental steps of the PCR reaction. The risk of infection of detection personnel and the risk of cross contamination between samples can be reduced to a certain extent based on the professional design of the PCR laboratory and the protection of personnel. However, the risk of personnel infection and cross-contamination of the sample is not completely avoided, since the sample tube and reagent tube still need to be repeatedly opened after sampling for sample and/or reagent transfer.
Disclosure of Invention
In order to overcome the problems in the prior art, the invention aims to provide a sampling tube for realizing nucleic acid amplification and detection without uncapping and an application method thereof.
The sampling tube capable of realizing nucleic acid amplification and detection without uncapping comprises a tube body and a tube cover, wherein a first protruding ring and a second protruding ring are arranged on the upper side of the outer wall of the tube body from top to bottom, a first concave ring and a second concave ring are arranged below the inner wall of the tube cover from top to bottom, the first protruding ring is matched with the first concave ring and the second concave ring and used for sealing the tube body and the tube cover, the second protruding ring is matched with the second concave ring and used for sealing the tube body and the tube cover, a waste liquid pool, a separation liquid storage structure and a contact pin structure are respectively arranged inside the tube cover from top to bottom, the contact pin structure is used for puncturing the separation liquid storage structure, a liquid draining channel is arranged between the waste liquid pool and the tube body, liquid is discharged from the tube body to the waste liquid pool when the tube body is inverted, and the liquid remains in the waste liquid pool when the tube body is placed vertically.
The separating liquid storage structure is provided with a separating ring and an outer ring, and is divided into an inner area and an outer area, and the bottom of the separating liquid storage structure is provided with a sealing film;
The contact pin structure comprises a wafer-shaped base and contact pins, the contact pins are fixed on the wafer-shaped base, when the pipe cover approaches to the pipe body, the wafer-shaped base takes the pipe orifice of the pipe body as a support, and the contact pins are used for piercing the sealing film.
The lengths of the contact pins corresponding to the inner area and the outer area are different, and the contact pins are used for releasing and separating liquid in the liquid storage structure step by step.
The sampling tube body is large in structure, small in size and conical in bottom, can be inserted into a reaction hole of a conventional PCR instrument, and has an opening end with a diameter of 1cm.
One end of the opening of the liquid discharge channel on the tube cover is positioned below the liquid storage separating structure, and the other end of the opening of the liquid discharge channel is positioned at the inner wall of the tube cover near the top of the cap, so that the reaction liquid is prevented from flowing back after entering the waste liquid chamber.
The cross sections of the convex parts of the first protruding ring and the second protruding ring are semicircular, the cross sections of the concave parts of the first concave ring and the second concave ring are semicircular, and the four parts have the same radius;
the center distance between the first protruding ring and the second protruding ring is the same as the center distance between the first concave ring and the second concave ring.
The separation ring and the outer ring are distributed in concentric circles, and the outer side face and the top face of the outer ring are connected with the inner wall and the top face of the pipe cover.
The 4 long hollow pins and 1 short hollow pin of the pin structure are distributed in a plum blossom shape on the disc-shaped base, wherein the 1 short hollow pin is positioned at the circle center;
The length difference value of the 4 long hollow pins and the short hollow pins is consistent with the center distance between the first protruding ring and the second protruding ring.
The edge of the wafer-shaped base of the contact pin structure is inwards recessed, the cross section of the wafer-shaped base is semicircular, and the wafer-shaped base is meshed with the sealing rubber ring.
According to the application method of the sampling tube, the inside of the tube body of the sampling tube is provided with a reaction reagent A, a reaction reagent B is arranged between the separation ring and the outer ring of the separation liquid storage structure, and a reaction reagent C is arranged in the separation ring;
The method comprises the steps of covering a tube cover after a sample is added into a sampling tube, adsorbing magnetic beads in a reaction reagent A on the bottom of the tube body through external magnetic force on the bottom of the tube body after reaction, rotating the whole sampling tube clockwise by 180 degrees to enable waste liquid to completely enter a waste liquid pool through a liquid discharge channel, rotating the sampling tube clockwise by 180 degrees, pressing the tube cover downwards until a first protruding ring is completely meshed with a second concave ring, penetrating a sealing film through a long hollow contact pin in a contact pin structure, releasing the reaction reagent B into the tube body, withdrawing an external magnet on the bottom of the tube body, rotating the tube body anticlockwise by 90 degrees, applying external magnetic force on a left tube wall to adsorb the magnetic beads, keeping the left tube wall continuously applying external magnetic force, rotating the tube body clockwise by 90 degrees, pressing the tube cover downwards until the first protruding ring is completely meshed with the first concave ring, the second protruding ring is completely meshed with the second concave ring, penetrating the sealing film through a short hollow contact pin in the contact pin structure, and releasing the reaction reagent C into the tube body for reaction.
Compared with the prior art, the method has the beneficial effects that through the technical scheme, after a sample is added, reagents required by the reaction can be added step by step under the condition of not uncovering, and the whole nucleic acid detection process is completed.
Drawings
FIG. 1 is a schematic view showing the structure of a sample tube in front view in an initial state for nucleic acid amplification and detection without opening the lid according to the present invention.
FIG. 2 is a schematic diagram showing the structure of a sample tube of the present invention in front view when a reagent B is added thereto without opening the cover to effect nucleic acid amplification and detection.
FIG. 3 is a schematic diagram showing the structure of a sample tube of the present invention in front view when a reagent C is added thereto without opening the cover to effect nucleic acid amplification and detection.
FIG. 4 is a schematic top view of a sample tube lancing device for nucleic acid amplification and detection without uncapping and a partial enlarged view of the engagement between the base and the sealing ring according to the present invention.
FIG. 5 is a schematic view showing the structure of a separating liquid storage structure in a tube cover of a sampling tube for amplifying and detecting nucleic acid without opening the cover.
FIG. 6 is a schematic top view of a sample tube with a cover in a cross section of a waste liquid chamber for nucleic acid amplification and detection without a cover.
In the figure, a tube body 10, a first protruding ring 11, a second protruding ring 12, a tube cover 20, a first concave ring 21, a second concave ring 22, a middle partition 23, a prism 24, a separating liquid storage structure 30, a separating ring 31, an outer ring 32, a sealing membrane 33, a pin structure 40, a wafer-shaped base 41, a long hollow pin 42, a short hollow pin 43 and a sealing rubber ring 44.
Detailed Description
In order that the invention may be readily understood, a more particular description of the invention will be rendered by reference to specific embodiments that are illustrated in the appended drawings and are described in connection with specific embodiments that are merely some, but not all, embodiments of the invention.
As shown in fig. 1 to 6, the invention provides a sampling tube for amplifying and detecting nucleic acid without opening a cover, which comprises a sampling tube body 10, wherein a tube cover 20 is arranged outside the top end of the sampling tube body, a separation liquid storage structure 30 is arranged inside the tube cover 20, and a contact pin structure 40 is arranged below the separation liquid storage structure 30.
The outer surface of the sampling tube body 10 near the opening end is provided with a first protruding ring 11 and a second protruding ring 12.
The lower part of the inner wall of the pipe cover 20 is provided with a first concave ring 21 and a second concave ring 22, the outer wall of the right side is provided with a raised prism 24, a liquid discharge channel is arranged in the prism 24, and a space between the inner wall of the top and the middle septum 23 is a waste liquid chamber.
The separate reservoir structure 30 includes a separate ring 31 and an outer ring 32 connected to the septum 23 inside the cap 20, and a sealing membrane 33 having a lower end for the needle insertion structure 40 to pierce.
The pin structure 40 comprises a disc-shaped base 41, 4 long hollow pins 42, 1 short hollow pin 43 and a sealing rubber ring 44.
Further, the sampling tube body 10 is large in structure, small in size and conical in bottom, and can be inserted into a reaction hole of a conventional PCR instrument, and the diameter of the near-opening end is 1cm.
Further, one end of the opening of the liquid draining channel on the tube cover 20 is located below the liquid separating and storing structure 30, and the other end is located at the inner wall of the tube cover 20 near the top of the cap, so as to prevent the reaction reagent from flowing back after entering the waste liquid chamber.
In this embodiment, after the tube cover 20 is tightly closed, the sampling tube is rotated clockwise by 180 ° to an inverted state, so that the reaction reagent a after the sample is processed enters the waste liquid chamber through the liquid discharge channel. Thereafter, the reaction reagent A is further rotated clockwise by 180 DEG to the initial position, and the reaction reagent A is prevented from flowing back from the waste liquid chamber.
Further, the tube cover 20 is engaged with the first concave ring 21 and the second concave ring 22 and the first protruding ring 11 and the second protruding ring 12 to control the relative position of the tube cover 20 and the sampling tube body 10. The cross sections of the convex parts of the first protruding ring 11 and the second protruding ring 12 are semicircular, the cross sections of the concave parts of the first concave ring 21 and the second concave ring 22 are semicircular, and the four have the same radius. The center distance between the first protruding ring 11 and the second protruding ring 12 is the same as the center distance between the first concave ring 21 and the second concave ring 22.
Further, the separation ring 31 and the outer ring 32 included in the separation liquid storage structure 30 are concentrically distributed, and the outer side surface and the middle septum of the outer ring 32 are connected with the inner wall and the middle septum of the tube cover 20.
Further, the 4 long hollow pins 42 and 1 short hollow pin 43 of the pin structure 40 are distributed in a quincuncial shape on the disc-shaped base 41, wherein the 1 short hollow pin 43 is located at a center of a circle, a round hole is formed in the disc-shaped base 41 at a position corresponding to the pin, so that the pin passes through the sealing film 33 and then the reaction reagent flows out, and a difference between lengths of the 4 long hollow pins 42 and the short hollow pin 43 is consistent with a center distance between the first protruding ring 11 and the second protruding ring 12. The edge of the disc-shaped base 41 of the pin structure 40 is recessed inwards, and the cross section of the disc-shaped base is semicircular and is meshed with the sealing rubber ring 44.
As shown in fig. 2, the long hollow pin 42 in the pin structure 40 pierces the sealing film 33 to release the reaction reagent B into the sample tube when the cap 20 is pressed to the first protruding ring 11 and the second recessed ring 22 to engage with each other, and the short hollow pin 43 in the pin structure 40 pierces the sealing film 33 to release the reaction reagent C into the sample tube when the cap 20 is continuously pressed to the first protruding ring 11 and the first recessed ring 21, the second protruding ring 12 and the second recessed ring 22 to engage with each other, thereby realizing the stepwise release of the reaction reagent B and the reaction reagent C.
Further, the inside of the sampling tube body 10 is provided with a reaction reagent A.
Further, a reactant B is disposed between the separation ring 31 and the outer ring 32 of the separation liquid storage structure 30, and a reactant C is disposed inside the separation ring 31.
The reaction reagent A/B/C may be charged with a suitable reagent according to the need and the reaction step.
Application example
A sampling tube for amplifying and detecting nucleic acid without opening the cover is prepared through covering the cover after the sample is added to the sampling tube. After a certain time of reaction, the magnetic beads in the reaction reagent A are adsorbed on the bottom of the tube body through magnetic force outside the bottom of the tube body, and the whole sampling tube is rotated 180 degrees clockwise, so that the waste liquid completely enters the waste liquid pool through the liquid discharge channel. And then the sampling tube is rotated 180 degrees clockwise, the tube cover is pressed downwards until the first protruding ring is completely meshed with the second concave ring, a long hollow contact pin in the contact pin structure pierces the sealing membrane, the reaction reagent B is released into the tube body, and the external magnet at the bottom of the tube body is withdrawn. After a certain time of reaction, the tube body is rotated anticlockwise for 90 degrees, external magnetic force is applied to the left tube wall to adsorb magnetic beads, the left tube wall is kept to continuously apply external magnetic force, the tube body is rotated clockwise for 90 degrees, the tube cover is pressed downwards until the first protruding ring is completely meshed with the first concave ring, the second protruding ring is completely meshed with the second concave ring, and a short hollow contact pin in the contact pin structure penetrates through the sealing film to release the reactant C into the tube body. After a certain time of reaction, the reaction result is observed by other equipment.
The foregoing is a further detailed description of the invention in connection with the preferred embodiments, and it is not intended that the invention be limited to the specific embodiments described. It will be apparent to those skilled in the art that several simple deductions or substitutions can be made without departing from the spirit of the invention, all of which fall within the scope of the invention as claimed.

Claims (9)

1.一种不开盖实现核酸扩增和检测的采样管，其特征在于，包括管体（10）、管盖（20）；1. A sampling tube for achieving nucleic acid amplification and detection without opening the cover, characterized in that it comprises a tube body (10) and a tube cover (20); 所述的管体（10）的外壁上侧从上到下设有第一突环（11）和第二突环（12）；The upper side of the outer wall of the tube body (10) is provided with a first protruding ring (11) and a second protruding ring (12) from top to bottom; 所述的管盖的内壁下方从上到下设有第一凹环（21）和第二凹环（22）；A first concave ring (21) and a second concave ring (22) are provided below the inner wall of the tube cover from top to bottom; 第一突环（11）和第一凹环（21）、第二凹环（22）相适配，用于密封管体（10）、管盖（20）；The first protruding ring (11) is matched with the first concave ring (21) and the second concave ring (22) and is used to seal the tube body (10) and the tube cover (20); 第二突环（12）和第二凹环（22）相适配，用于密封管体（10）、管盖（20）；The second protruding ring (12) and the second concave ring (22) are matched to each other and are used to seal the tube body (10) and the tube cover (20); 所述管盖（20）内部从上到下分别设有废液池、分隔储液结构（30）、插针结构（40）；The tube cover (20) is provided with a waste liquid pool, a partition liquid storage structure (30), and a pin insertion structure (40) from top to bottom. 插针结构（40）用于刺破分隔储液结构（30）；所述插针结构（40）包括圆片形底座（41）和插针，插针固定在圆片形底座（41）上，当管盖（20）接近管体（10）时，圆片形底座（41）以管体（10）的管口为支撑；The insertion needle structure (40) is used to puncture the partition liquid storage structure (30); the insertion needle structure (40) comprises a disc-shaped base (41) and an insertion needle, the insertion needle is fixed on the disc-shaped base (41), and when the tube cover (20) approaches the tube body (10), the disc-shaped base (41) is supported by the tube mouth of the tube body (10); 所述管盖（20）上的排液通道的开口一端位于分隔储液结构（30）下方，另一端位于所述管盖（20）近帽顶内壁处，以防止反应液进入废液室后反流；One end of the opening of the liquid discharge channel on the tube cover (20) is located below the partition liquid storage structure (30), and the other end is located near the inner wall of the cap top of the tube cover (20) to prevent the reaction liquid from flowing back after entering the waste liquid chamber; 废液池和管体（10）之间设有排液通道，倒置时，液体从管体（10）排到废液池，正置时，液体留存在废液池。A liquid discharge channel is provided between the waste liquid pool and the tube body (10); when the tube body (10) is inverted, the liquid is discharged from the tube body (10) into the waste liquid pool; when the tube body (10) is upright, the liquid remains in the waste liquid pool. 2.根据权利要求1所述的采样管，其特征在于，所述的分隔储液结构（30）设有分隔环（31）和外环（32），分为内外两区，分隔储液结构（30）的底部设有密封膜（33）。2. The sampling tube according to claim 1 is characterized in that the partition liquid storage structure (30) is provided with a partition ring (31) and an outer ring (32), which is divided into inner and outer areas, and a sealing film (33) is provided at the bottom of the partition liquid storage structure (30). 3.根据权利要求2所述的采样管，其特征在于，所述的内外两区对应的插针长度不同，用于分步释放分隔储液结构（30）内的液体。3. The sampling tube according to claim 2, characterized in that the lengths of the insertion pins corresponding to the inner and outer areas are different, and are used to release the liquid in the separated liquid storage structure (30) in steps. 4.根据权利要求1所述的采样管，其特征在于，所述采样管管体（10）结构上大下小，底部呈圆锥形，可插入常规PCR仪反应孔，开口端直径为1cm。4. The sampling tube according to claim 1 is characterized in that the sampling tube body (10) is larger at the top and smaller at the bottom, and the bottom is conical, which can be inserted into the reaction hole of a conventional PCR instrument, and the diameter of the open end is 1 cm. 5.根据权利要求1所述的采样管，其特征在于，所述第一突环（11）和所述第二突环（12）的凸起部分横截面为半圆形，所述第一凹环（21）和所述第二凹环（22）的凹陷部分横截面为半圆形，四者具有相同的半径；5. The sampling tube according to claim 1, characterized in that the cross-section of the raised portion of the first protruding ring (11) and the second protruding ring (12) is semicircular, the cross-section of the recessed portion of the first concave ring (21) and the second concave ring (22) is semicircular, and the four have the same radius; 所述第一突环（11）和所述第二突环（12）的圆心间距与所述第一凹环（21）和所述第二凹环（22）的圆心间距相同。The distance between the centers of the first protruding ring (11) and the second protruding ring (12) is the same as the distance between the centers of the first concave ring (21) and the second concave ring (22). 6.根据权利要求2所述的采样管，其特征在于，所述分隔环（31）和所述外环（32）呈同心圆分布，所述外环（32）的外侧面与顶面均与所述管盖（20）的内壁和顶面相连。6. The sampling tube according to claim 2, characterized in that the separation ring (31) and the outer ring (32) are distributed in concentric circles, and the outer side surface and the top surface of the outer ring (32) are connected to the inner wall and the top surface of the tube cover (20). 7.根据权利要求2所述的采样管，其特征在于，所述插针结构（40）的4根长空心插针（42）和1根短空心插针（43）在所述圆片形底座（41）上呈梅花形分布，其中所述1根短空心插针（43）位于圆心；所述圆片形底座（41）对应插针的位置设有圆孔，以便于插针穿过所述密封膜（33）后反应试剂流出；7. The sampling tube according to claim 2, characterized in that the four long hollow pins (42) and one short hollow pin (43) of the pin structure (40) are distributed in a plum blossom shape on the disc-shaped base (41), wherein the one short hollow pin (43) is located at the center of the circle; a circular hole is provided in the disc-shaped base (41) at a position corresponding to the pin, so that the reaction reagent can flow out after the pin passes through the sealing film (33); 所述4根长空心插针（42）和所述短空心插针（43）的长度差值与所述第一突环（11）和所述第二突环（12）的圆心间距一致。The difference in length between the four long hollow pins (42) and the short hollow pin (43) is consistent with the distance between the centers of the first protruding ring (11) and the second protruding ring (12). 8.根据权利要求1所述的采样管，其特征在于，所述插针结构（40）的所述圆片形底座（41）边缘向内凹陷，截面呈半圆形，与密封胶圈（44）啮合。8. The sampling tube according to claim 1, characterized in that the edge of the disc-shaped base (41) of the pin structure (40) is concave inwards, has a semicircular cross-section, and is engaged with the sealing rubber ring (44). 9.一种根据权利要求7所述的采样管的应用方法，其特征在于，所述采样管管体（10）内部装有反应试剂A；所述分隔储液结构（30）的所述分隔环（31）和所述外环（32）之间装有反应试剂B，所述分隔环（31）内部装有反应试剂C；9. An application method of the sampling tube according to claim 7, characterized in that the sampling tube body (10) is filled with a reaction reagent A; a reaction reagent B is filled between the separation ring (31) and the outer ring (32) of the separation liquid storage structure (30), and a reaction reagent C is filled inside the separation ring (31); 步骤如下：当样品被加入采样管后，盖上管盖（20），在反应后，通过在管体（10）底部外部磁力将反应试剂A中的磁珠吸附在管体（10）底部，将采样管整体顺时针旋转180°，使废液通过排液通道完全进入废液池，再将采样管顺时针旋转180°，向下按压管盖（20）至第一突环（11）与第二凹环（22）完全啮合，插针结构（40）中的长空心插针（42）穿刺密封膜（33），将反应试剂B释放入管体中，撤销管体（10）底部的外部磁铁；在反应后，将管体逆时针旋转90°，并在左侧管壁施加外部磁力吸附磁珠；保持左侧管壁持续施加外部磁力并将管体顺时针旋转90°，向下按压管盖（20）至第一突环（11）与第一凹环（21）、第二突环（12）与第二凹环（22）完全啮合，插针结构（40）中的短空心插针（43）穿刺密封膜（33），将反应试剂C释放入管体中进行反应。The steps are as follows: after the sample is added to the sampling tube, the tube cover (20) is covered. After the reaction, the magnetic beads in the reaction reagent A are adsorbed on the bottom of the tube body (10) by the external magnetic force at the bottom of the tube body (10). The sampling tube is rotated 180 degrees clockwise as a whole, so that the waste liquid completely enters the waste liquid pool through the drainage channel. The sampling tube is then rotated 180 degrees clockwise, and the tube cover (20) is pressed downward until the first protruding ring (11) and the second concave ring (22) are completely engaged. The long hollow needle (42) in the needle structure (40) pierces the sealing film (33), and the reaction reagent A is discharged. B is released into the tube body, and the external magnet at the bottom of the tube body (10) is removed; after the reaction, the tube body is rotated 90° counterclockwise, and an external magnetic force is applied to the left tube wall to adsorb the magnetic beads; the left tube wall is kept continuously applying the external magnetic force and the tube body is rotated 90° clockwise, and the tube cover (20) is pressed downward until the first protruding ring (11) and the first concave ring (21), and the second protruding ring (12) and the second concave ring (22) are fully engaged, and the short hollow pin (43) in the pin structure (40) pierces the sealing film (33), and the reaction reagent C is released into the tube body for reaction.
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