A storage and addition device for biological sample pretreatment additives

Abstract

This application relates to the field of pretreatment of biological samples, and particularly to a storage and addition device for additives for pretreatment of biological samples. The device includes a cover body and a piercing device. The cover body includes a spiral tube, a storage tube, and a pressing stroke tube. The inside of the spiral tube is provided with threads. The pressing stroke tube is located above the spiral tube. The storage tube is arranged in the inner cavity of the cover body and is integrally connected to the cover body through a connecting part. The bottom end of the storage tube is closed. The piercing device includes a piercing rod, a pressing head, and a sealing rubber piston. The piercing rod is connected to the pressing head. The lower part of the piercing rod is located in the storage tube. The pressing head is arranged on the pressing stroke tube. The sealing rubber piston is fixedly arranged on the piercing rod. The outer wall of the sealing rubber piston fits with the inner wall of the storage tube. Pressing down the pressing head can make the piercing rod pierce through the bottom of the storage tube, and the pretreatment additive in the storage tube can be pushed out from the bottom of the storage tube by the sealing rubber piston. This application solves the technical problem that after piercing through the bottom of the storage tube, due to air pressure difference or surface tension, the additive in the storage tube may not completely fall into the centrifuge column, and the addition is convenient and fast.

Description

Technical Field

[0001] This application relates to the field of biological sample pretreatment, and particularly to a device for storing and adding biological sample pretreatment additives. Background Art

[0002] Real-time fluorescence quantitative PCR technology has been widely used in many fields such as molecular diagnosis of genetic diseases, clinical examination, quarantine of imported and exported animals and plants, food safety monitoring, soil microorganism detection, and paternity testing. Since samples such as blood, food, and soil contain a large number of inhibitory factors, such as hemoglobin, hemin, lactoferrin, and humic acid, they will have an obvious inhibitory effect on conventional Taq DNA polymerase. Therefore, nucleic acids must be first isolated and extracted from these samples to be tested and then used for PCR amplification. Nucleic acid extraction is the first step in nucleic acid detection and one of the key methods in molecular biology. It provides a basis for downstream nucleic acid detection, and the extraction quality and integrity directly affect clinical research or diagnosis.

[0003] Generally, the nucleic acid extraction process includes three steps: sample lysis, binding, and purification. Traditional extraction reagents have relatively low yield rates and complicated steps, which have a certain impact on the extraction work. Especially when the sample volume is scarce, it is difficult to obtain qualified nucleic acids. Moreover, traditional methods have many operation steps, high costs, large sample requirements, and are prone to cross-contamination, which is not conducive to customers quickly extracting and purifying nucleic acid samples.

[0004] In addition, in the currently most widely used real-time fluorescence PCR technology for nucleic acid detection, most of the reagents are not ready-to-use, resulting in problems such as complicated operation, time-consuming, and high costs in PCR detection. Before use, complex configuration work is required, which is likely to cause experimental errors or even failures. Different personnel operations also cause experimental errors; it is impossible to achieve one-step rapid nucleic acid extraction; PCR detection reagents need to be manually configured on-site, multiple steps require manual marking, and the detection instruments at each stage are independent, which is not conducive to fully automatic processing of a large number of test samples. Summary of the Invention

[0005] In order to solve the above technical problems, the purpose of this application is to provide a device for storing and adding biological sample pretreatment additives. This device can be provided with additives in a storage tube. By pressing down the pressing head, the piercing rod can pierce through the bottom of the storage tube, and the pretreatment additive in the storage tube can be pushed out from the bottom of the storage tube through the sealing rubber piston, avoiding on-site configuration work and making the addition convenient and fast.

[0006] In order to achieve the above purpose, this application adopts the following technical solutions:

[0007] A storage and addition device for a biological sample pretreatment additive. The device includes a cover body and a piercing device. The cover body includes a spiral tube, a storage tube, and a pressing stroke tube. The inside of the spiral tube is provided with threads. The pressing stroke tube is located above the spiral tube. The storage tube is arranged in the inner cavity of the cover body and is integrally connected to the cover body through a connecting part. The bottom end of the storage tube is closed. The piercing device includes a piercing rod, a pressing head, and a sealing rubber piston. The piercing rod is connected to the pressing head. The lower part of the piercing rod is located in the storage tube. The pressing head is arranged on the pressing stroke tube. The sealing rubber piston is fixedly arranged on the piercing rod. The outer wall of the sealing rubber piston fits with the inner wall of the storage tube. Pressing down the pressing head can cause the piercing rod to pierce through the bottom of the storage tube, and the pretreatment additive in the storage tube can be pushed out from the bottom of the storage tube through the sealing rubber piston.

[0008] As a further improvement, the upper part of the piercing rod is a guiding rod, and the guiding rod is adapted to the inner wall of the storage tube. The upper end of the guiding rod is fixedly connected to the pressing head.

[0009] In this application, during storage and transportation, due to misoperation or air pressure difference problems, the pressing head may be pressed down, resulting in the piercing of the bottom of the storage tube. Therefore, as an improvement, in this application, the pressing head is provided with a downward annular sleeve. The lower part of the annular sleeve is provided with a convex point protruding from the annular sleeve, and a notch adapted to the convex point is arranged on the inner side of the upper part of the pressing stroke tube. The convex point is clamped in the notch to form a positioning. In this way, a limit is formed, and the pressing head will not be pressed down due to misoperation when there is no normal operation.

[0010] As a further improvement, strip-shaped grooves are respectively arranged on both sides of the convex point. The two strip-shaped grooves make the convex point located on an elastic piece, which has good elasticity and is convenient for the pressing head to press down to break through the limit.

[0011] As a further improvement, the bottom of the piercing rod is a pointed head, and a liquid guiding groove is axially arranged above the pointed head. The liquid guiding groove can prevent the piercing rod from being closely attached to the damaged position at the bottom, resulting in the blockage of the additive from falling.

[0012] As a further improvement, the additive is one or more of a lysis solution, a purification solution, a protein adsorption material, and a protein precipitation material. If the centrifuge tube contains a preservation solution, then the additive can be a lysis solution; if the centrifuge tube contains a lysis solution, then the additive can be one or more of a purification solution, a protein adsorption material, and a protein precipitation material, so that lysis and purification can be completed in one step. Since samples such as blood, food, and soil contain a large amount of hemoglobin, hemin, lactoferrin, humic acid, etc., therefore, more preferably, the additive is a protein adsorption material and a protein precipitation material; especially, most preferably, the additive is a hemoglobin adsorption material, which can avoid the influence of hemoglobin on fluorescence PCR.

[0013] As a further improvement, the bottom sealing material of the storage tube is aluminum foil or tin foil. Of course, it can also be to seal the bottom of the storage tube by thinning the bottom sheet.

[0014] Since the present application adopts the above technical solution, it solves the technical problem that after piercing the bottom of the storage tube, due to air pressure difference or surface tension, the additives in the storage tube may not completely fall into the centrifuge column, and the addition is convenient and fast. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 It is a schematic structural diagram of the present application.

[0016] Figure 2 It is an exploded structural diagram of the present application.

[0017] Figure 3 It is a schematic structural diagram of a centrifuge tube.

[0018] Figure 4 It is a schematic structural diagram of the test tube cap of Example 2.

[0019] Figure 5 It is a schematic structural diagram of the test tube cap of Example 3. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0020] The following will describe in detail the specific embodiments of the present application with reference to the accompanying drawings.

[0021] Example 1

[0022] As Figure 1 shown, a pretreatment device for purifying trace biological samples includes a collection tube 1, a centrifuge column 2, a test tube cap 7 and a reagent solution 3. The test tube cap 7 is threadedly connected to the collection tube 1. The centrifuge column 2 is disposed in the collection tube 1. A convex ring 6 is provided at the upper part of the centrifuge column 2. The centrifuge column 2 is disposed on the upper end surface of the collection tube 1 through the convex ring 6, and a liquid collection cavity is formed at the lower part of the centrifuge column 2. The centrifuge column 2 is hung on the collection tube 1 by being disposed on the upper end surface of the collection tube 1 through the convex ring 6, and the centrifuge column 2 can be locked by setting the test tube cap 7.

[0023] As Figure 1 shown, the test tube cap 7 is disposed at the upper end of the collection tube 1 to seal the collection tube 1. The test tube cap 7 includes a cap body and a piercing device. The cap body includes a spiral tube, a storage tube 8 and a pressing travel tube. Threads are provided inside the spiral tube. The pressing travel tube is located at the upper part of the spiral tube. The storage tube 8 is disposed in the inner cavity of the cap body and is integrally connected to the cap body through a connecting portion. The bottom end of the storage tube 8 is sealed; As Figure 2As shown, the pressing head 13 is provided with a downward annular sleeve. The lower part of the annular sleeve is provided with bumps 21 protruding from the annular sleeve, and notches 22 adapted to the bumps 21 are provided on the inner side of the upper part of the pressing stroke tube. The bumps 21 are clamped in the notches 22 to form positioning, and strip-shaped grooves 23 are respectively provided on both sides of the bumps 21.

[0024] As Figure 1 shown, the piercing device includes a piercing rod 12, a pressing head 13 and a sealing rubber piston 14. The upper part of the piercing rod 12 is a guiding rod 11, which is adapted to the inner wall of the storage tube 8. The upper end of the guiding rod 11 is fixedly connected to the pressing head 13. The lower part of the piercing rod 12 is located inside the storage tube 8. The bottom of the piercing rod 12 is a pointed head, and a liquid guiding groove 24 is axially provided above the pointed head. The pressing head 13 is arranged on the pressing stroke tube. The sealing rubber piston 14 is fixedly arranged on the piercing rod 12, and the outer wall of the sealing rubber piston 14 fits with the inner wall of the storage tube 8. Pressing down the pressing head 13 can cause the piercing rod 12 to pierce through the bottom of the storage tube 8, and the pretreatment additive in the storage tube 8 can be pushed out from the bottom of the storage tube 8 through the sealing rubber piston 14 and fall into the centrifuge column 2.

[0025] As Figure 3 shown, a surface tension layer 4 is provided at the bottom of the centrifuge column 2, and a common filter layer 5 is provided above the surface tension layer 4. The surface tension layer 4 adopts a hydrophobic sieve plate, which is sintered from ultra-high molecular weight polyethylene (UHMW-PE). The filtration accuracy of the hydrophobic sieve plate is 10-100 microns. The reagent solution 3 is arranged inside the centrifuge column 2 and above the surface tension layer 4. The liquid of the reagent solution 3 has surface tension on the surface tension layer 4 to overcome its own gravity so that the liquid cannot pass through the surface tension layer 4. When conducting an experiment, the whole device can be placed in a centrifuge. When the centrifuge column 2 is under the action of centrifugal force, the liquid overcomes the surface tension and drops from the centrifuge column 2 into the collection tube 1. Alternatively, the test tube cap 7 can be opened, and the liquid can be pushed into the collection tube 1 by setting an injection piston rod. Of course, when ensuring sufficient sealing between the test tube cap 7, the centrifuge column 2 and the collection tube 1, the liquid can also be pushed into the collection tube 1 through the upper sealing rubber piston 14 of the piercing device.

[0026] Taking the extraction of nucleic acid from blood as an example, the reagent solution 3 in this embodiment is a lysis solution, and the additive 9 is a hemoglobin adsorption material.

[0027] The nucleic acid purification pretreatment method of this embodiment includes the following steps:

[0028] 1) Add the reagent solution 3 into the centrifuge column 2, or the centrifuge column 2 itself is preset with the reagent solution 3;

[0029] 2) Add the sample to be extracted into the centrifuge column 2, and lyse the sample to be extracted under the action of the reagent solution 3;

[0030] 3) Add or not add the additive 9;

[0031] 4) Place the centrifuge column 2 collection tube 1 in a centrifuge and centrifuge. Under the action of centrifugal force, the liquid containing nucleic acid overcomes the surface tension and drips from the centrifuge column 2 into the collection tube 1.

[0032] Example 2

[0033] As Figure 4 shown, the piercing device includes a piercing rod 12 and a pressing head 15. The pressing head 15 is a rubber ball head or a plastic folding head. An opening is provided in the middle of the test tube cap 7, and the storage tube 8 is arranged in the inner cavity of the test tube cap 7 and connected into one body through a connecting part; the pressing head 15 is located above the storage tube 8, and the upper part of the piercing rod 12 is fixedly connected to the inner upper wall of the pressing head 15. By pressing the pressing head 15, the piercing rod 12 presses down to pierce the bottom of the storage tube 8, and at the same time, the additive 9 is pushed into the centrifuge column 2 through the air pressure change of the pressing head 15. Other structures of this embodiment are as shown in Example 1.

[0034] Example 3

[0035] As Figure 5 shown, the piercing device includes a piercing rod 12, a rotating body 16 and a guiding platform 17. An opening is provided in the middle of the test tube cap 7, and the storage tube 8 is fixedly arranged below the opening. The rotating body 16 is located on the opening, and the rotating body 16 is rotationally matched with the opening. The guiding platform 17 is arranged in the rotating body 16, and the guiding platform 17 is threadedly connected with the inner wall of the rotating body 16. An anti-rotation guiding rib 19 and a groove 18 are provided between the lower part of the guiding platform 17 and the opening. The piercing rod 12 is arranged below the guiding platform 17. By rotating the rotating body 16, the guiding platform 17 is driven to move downward, and the piercing rod 12 presses down to pierce the bottom of the storage tube 8. A ventilation channel 20 can be arranged inside or on the side wall of the guiding platform 17, and the additive falls into the centrifuge column 2 by the action of gravity. Other structures of this embodiment are as shown in Example 1.

[0036] The above is the description of the embodiments of the present invention. Through the above description of the disclosed embodiments, those skilled in the art can implement or use the present invention. Various modifications to these embodiments will be obvious to those skilled in the art. The general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but will be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. An integrated adding device for biological sample pretreatment reagents, characterized in that, the device includes a collection tube (1), a centrifuge column (2), a test tube cap (7), a reagent solution (3) and a pretreatment additive (9). The centrifuge column (2) is arranged inside the collection tube (1), and the reagent solution (3) is arranged inside the centrifuge column (2); the test tube cap (7) is arranged at the upper end of the collection tube (1), and the test tube cap (7) includes a cap body and a piercing device. The cap body includes a spiral tube, a storage tube (8) and a pressing stroke tube. Threads are arranged inside the spiral tube, and the pressing stroke tube is located above the spiral tube. The storage tube (8) is arranged inside the cavity of the cap body and is integrally connected with the cap body through a connecting part. The bottom end of the storage tube (8) is closed; the piercing device includes a piercing rod (12), a pressing head (13) and a sealing rubber piston (14). The piercing rod (12) is connected with the pressing head (13). The lower part of the piercing rod (12) is located inside the storage tube (8). The pressing head (13) is arranged on the pressing stroke tube. The sealing rubber piston (14) is fixedly arranged on the piercing rod (12), and the outer wall of the sealing rubber piston (14) fits with the inner wall of the storage tube (8). Pressing down the pressing head (13) can make the piercing rod (12) pierce through the bottom of the storage tube (8), and the pretreatment additive (9) inside the storage tube (8) is pushed out from the bottom of the storage tube (8) through the sealing rubber piston (14); after the bottom of the storage tube (8) is pierced, the pretreatment additive (9) inside the storage tube (8) falls into the centrifuge column (2); the reagent solution (3) is a preservation solution, and the pretreatment additive (9) is a lysis solution; or, the reagent solution (3) is a lysis solution, and the pretreatment additive (9) is one or more of a purification solution, a protein adsorption material and a protein precipitation material.

2. The device according to claim 1, characterized in that, the upper part of the piercing rod (12) is a guiding rod (11), and the guiding rod (11) is adapted to the inner wall of the storage tube (8). The upper end of the guiding rod (11) is fixedly connected with the pressing head (13).

3. The device according to claim 1, characterized in that, the pressing head (13) is provided with a downward annular sleeve, and convex points (21) protruding from the annular sleeve are arranged at the lower part of the annular sleeve. A notch (22) adapted to the convex points (21) is arranged on the inner side of the upper part of the pressing stroke tube. The convex points (21) are clamped in the notch (22) to form positioning.

4. The device according to claim 3, characterized in that, strip-shaped grooves (23) are respectively arranged on both sides of the convex points (21).

5. The device according to claim 1, characterized in that, the pretreatment additive (9) is a hemoglobin adsorption material.

6. The device according to claim 1, characterized in that, the bottom closing material (10) of the storage tube (8) is aluminum foil or tin foil.

7. The device according to claim 1, characterized in that, the bottom of the piercing rod (12) is a pointed head, and a liquid guiding groove (24) is axially arranged above the pointed head.

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