CN111286448A

CN111286448A - Biological sample pretreatment additive storage and addition device

Info

Publication number: CN111286448A
Application number: CN202010251534.7A
Authority: CN
Inventors: 王德明; 周杰锋
Original assignee: Ningbo Ajcore Biotechnology Co ltd
Current assignee: Ningbo Ajcore Biotechnology Co ltd
Priority date: 2020-04-01
Filing date: 2020-04-01
Publication date: 2020-06-16

Abstract

The application relates to the field of biological sample pretreatment, in particular to a device for storing and adding a biological sample pretreatment additive. The device comprises a cover body and a puncturing device, wherein the cover body comprises a spiral pipe, a storage pipe and a pressing stroke pipe, threads are arranged in the spiral pipe, the pressing stroke pipe is positioned at the upper part of the spiral pipe, the storage pipe is arranged in an inner cavity of the cover body and is connected with the cover body into a whole through a connecting part, and the bottom end of the storage pipe is closed; the breaking device comprises a breaking rod, a pressing head and a sealing rubber piston, the breaking rod is connected with the pressing head, the lower portion of the breaking rod is located in the storage pipe, the pressing head is arranged on the pressing stroke pipe, the sealing rubber piston is fixedly arranged on the breaking rod, the outer wall of the sealing rubber piston is attached to the inner wall of the storage pipe, the pressing head is pressed downwards to enable the breaking rod to break the bottom of the storage pipe, and pretreatment additives in the storage pipe are pushed to flow out of the bottom of the storage pipe through the sealing rubber piston. The technical problem that the additive in the storage pipe can not completely fall into the centrifugal column due to air pressure difference or surface tension after the bottom of the storage pipe is broken through is solved, and the additive is convenient and fast to add.

Description

Biological sample pretreatment additive storage and addition device

Technical Field

The application relates to the field of biological sample pretreatment, in particular to a device for storing and adding a biological sample pretreatment additive.

Background

The real-time fluorescent quantitative PCR technology is widely applied to a plurality of fields such as genetic disease molecular diagnosis, clinical examination, animal and plant import and export quarantine, food safety monitoring, soil microorganism detection, paternity test and the like. Because samples such as blood, food, soil and the like contain a large amount of inhibiting factors such as hemoglobin, methemoglobin, lactoferrin, humic acid and the like, the Taq DNA polymerase has obvious inhibiting effect on the conventional Taq DNA polymerase. Therefore, nucleic acids must be isolated from these test samples and then used for PCR amplification. Nucleic acid extraction is the first step of nucleic acid detection, and is also one of the key methods in molecular biology. Provides a basis for downstream nucleic acid detection, and the quality and integrity of extraction directly influence clinical research or diagnosis.

The general nucleic acid extraction process comprises three steps: lysis, binding and purification of the sample. The traditional extraction reagent has relatively low extraction rate and complex steps, thus bringing certain influence on extraction work, and particularly being difficult to obtain qualified nucleic acid when the sample amount is rare. In addition, the traditional method has the disadvantages of multiple operation steps, high cost, large sample requirement, easy cross contamination and inconvenience for customers to quickly extract and purify nucleic acid samples.

In addition, in the current real-time fluorescence PCR technology with the widest application of nucleic acid detection, most reagents are not ready-to-use, so that the problems of complicated operation, high labor and cost and the like of PCR detection generally exist, complicated configuration work is required before use, and errors and even failures of tests are easily caused. Experimental errors also arise from different personnel operations; the rapid extraction of nucleic acid by one-step method cannot be realized; PCR detection reagents need to be manually configured on site, multiple steps need to be manually marked, detection instruments at all stages are independent, and full-automatic treatment of a large number of test samples is not facilitated.

Disclosure of Invention

In order to solve the above technical problems, an object of the present application is to provide a device for storing and adding a pretreatment additive in a biological sample, which can be used for arranging an additive in a storage tube, wherein a breaking rod can break the bottom of the storage tube by pressing a pressing head downwards, and a rubber sealing piston pushes the pretreatment additive in the storage tube to flow out from the bottom of the storage tube, thereby avoiding field configuration work and facilitating and rapid addition.

In order to achieve the above object, the present application adopts the following technical solutions:

a biological sample pretreatment additive storage and addition device comprises a cover body and a puncturing device, wherein the cover body comprises a spiral pipe, a storage pipe and a pressing stroke pipe, threads are arranged in the spiral pipe, the pressing stroke pipe is positioned at the upper part of the spiral pipe, the storage pipe is arranged in an inner cavity of the cover body and is connected with the cover body into a whole through a connecting part, and the bottom end of the storage pipe is closed; the breaking device comprises a breaking rod, a pressing head and a sealing rubber piston, the breaking rod is connected with the pressing head, the lower portion of the breaking rod is located in the storage pipe, the pressing head is arranged on the pressing stroke pipe, the sealing rubber piston is fixedly arranged on the breaking rod, the outer wall of the sealing rubber piston is attached to the inner wall of the storage pipe, the pressing head is pressed downwards to enable the breaking rod to break the bottom of the storage pipe, and pretreatment additives in the storage pipe are pushed to flow out of the bottom of the storage pipe through the sealing rubber piston.

As a further improvement, the upper part of the breaking rod is a guide rod, the guide rod is matched with the inner wall of the storage pipe, and the upper end of the guide rod is fixedly connected with the pressing head.

In this application, when storing the transportation, because maloperation or the poor problem of atmospheric pressure, the press head can push down and lead to the storage tube bottom to wear brokenly, for this reason, as an improvement, this application, the press head be provided with decurrent annular cover, the lower part of annular cover is provided with the bump of protrusion annular cover to the upper portion inboard of pressing the stroke pipe be provided with the notch of bump looks adaptation, the bump joint forms the location in the notch. Therefore, a limit is formed, and the pressing head cannot be pressed down due to misoperation when normal operation is not performed.

As further improvement, bump both sides be provided with the bar groove respectively, two bar grooves make the bump be located an elastic sheet, have better elasticity like this, the press head of being convenient for pushes down and breaks through spacingly.

As a further improvement, the bottom of the breaking rod is a sharp head, a liquid guide groove is axially arranged above the sharp head, and the phenomenon that the breaking rod is tightly attached to the bottom broken position to cause the falling of the additive to be blocked can not occur in the liquid guide groove.

As a further improvement, the additive is one or more of lysis solution, purification solution, protein adsorption material and protein precipitation material. If the centrifuge tube is a preservation solution, the additive can be a lysis solution; if the centrifuge tube contains a lysis solution, the additive may be one or more of a purification solution, a protein adsorbent material and a protein precipitation material, so that lysis and purification are completed in one step. Because samples such as blood, food, soil and the like contain a large amount of hemoglobin, methemoglobin, lactoferrin, humic acid and the like, the additives are preferably protein adsorbing materials and protein precipitating materials; particularly, the additive is most preferably hemoglobin adsorbing material, so that the influence of hemoglobin on the fluorescence PCR can be avoided.

As a further improvement, the bottom sealing material of the storage tube is aluminum foil or tin foil. Of course, the bottom of the storage tube may be flaked and sealed.

By adopting the technical scheme, the technical problem that the additive in the storage pipe can not completely fall into the centrifugal column due to air pressure difference or surface tension after the bottom of the storage pipe is broken through is solved, and the addition is convenient and fast.

Drawings

Fig. 1 is a schematic structural diagram of the present application.

Fig. 2 is a schematic diagram of the explosive structure of the present application.

FIG. 3 is a schematic view of the structure of the centrifuge tube.

FIG. 4 is a schematic view showing the structure of a test tube cover in example 2.

FIG. 5 is a schematic view showing the structure of a lid for a test tube in example 3.

Detailed Description

The following detailed description of embodiments of the present application will be made with reference to the accompanying drawings.

Example 1

The device comprises a collecting pipe 1, a centrifugal column 2, a test tube cover 7 and a reagent solution 3. The centrifugal column is characterized in that the test tube cover 7 is connected with the collecting pipe 1 through threads, the centrifugal column 2 is arranged in the collecting pipe 1, a convex ring 6 is arranged on the upper portion of the centrifugal column 2, the centrifugal column 2 is arranged on the upper end face of the collecting pipe 1 through the convex ring 6, and a liquid collecting cavity is formed in the lower portion of the centrifugal column 2. The centrifugal column 2 is arranged on the upper end surface of the collecting pipe 1 through a convex ring 6, so that the centrifugal column 2 is hung on the collecting pipe 1, and the centrifugal column 2 can be locked by arranging a test tube cover 7.

As shown in fig. 1, the test tube cover 7 is arranged at the upper end of the collecting tube 1 to seal the collecting tube 1, the test tube cover 7 comprises a cover body and a puncturing device, the cover body comprises a spiral tube, a storage tube 8 and a pressing stroke tube, threads are arranged in the spiral tube, the pressing stroke tube is positioned at the upper part of the spiral tube, the storage tube 8 is arranged in the inner cavity of the cover body and is connected with the cover body into a whole through a connecting part, and the bottom end of the storage tube 8 is sealed; as shown in fig. 2, the pressing head 13 is provided with a downward annular sleeve, the lower part of the annular sleeve is provided with a protruding point 21 protruding out of the annular sleeve, the inner side of the upper part of the pressing stroke tube is provided with a notch 22 matched with the protruding point 21, the protruding point 21 is clamped in the notch 22 to form a positioning, and two sides of the protruding point 21 are respectively provided with a strip-shaped groove 23.

As shown in fig. 1, the puncturing device comprises a puncturing rod 12, a pressing head 13 and a sealing rubber piston 14, wherein the upper part of the puncturing rod 12 is a guide rod 11, the guide rod 11 is matched with the inner wall of the storage tube 8, and the upper end of the guide rod 11 is fixedly connected with the pressing head 13. The lower part of the breaking-through rod 12 is positioned in the storage pipe 8, the bottom of the breaking-through rod 12 is a tip, and a liquid guide groove 24 is axially arranged above the tip. The pressing head 13 is arranged on the pressing stroke pipe, the sealing rubber piston 14 is fixedly arranged on the piercing rod 12, the outer wall of the sealing rubber piston 14 is attached to the inner wall of the storage pipe 8, the piercing rod 12 can pierce the bottom of the storage pipe 8 by pressing the pressing head 13 downwards, and the sealing rubber piston 14 pushes the pretreatment additive in the storage pipe 8 to flow out of the bottom of the storage pipe 8 and fall into the centrifugal column 2.

As shown in FIG. 3, the bottom of the centrifugal column 2 is provided with a surface tension layer 4, and the upper part of the surface tension layer 4 is provided with a common filter layer 5. The surface tension layer 4 is made of a hydrophobic sieve plate which is formed by sintering ultra-high molecular weight polyethylene (UHMW-PE), and the filtering precision of the hydrophobic sieve plate is 10-100 microns. The reagent solution 3 is arranged in the centrifugal column 2 and is positioned above the surface tension layer 4, and the liquid of the reagent solution 3 has surface tension on the surface tension layer 4 so as to overcome the gravity of the liquid and prevent the liquid from passing through the surface tension layer 4. When the experiment is carried out, the whole set of device can be placed in a centrifuge, and when the centrifugal column 2 is acted by centrifugal force, liquid overcomes surface tension and drops into the collecting pipe 1 from the centrifugal column 2. Alternatively, the vial cap 7 may be opened and the liquid pushed into the collection tube 1 by providing an injection plunger rod. Of course, it is also possible to push liquid into the collection tube 1 by breaking the rubber sealing piston 14 at the upper end of the device, while ensuring a sufficient seal between the cuvette lid 7, the spin column 2 and the collection tube 1.

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-adsorbing material.

The method for pretreatment of nucleic acid for purification of the present embodiment comprises the steps of:

1) adding the reagent solution 3 into the centrifugal column 2, or presetting the reagent solution 3 in the centrifugal column 2;

2) adding the extracted sample into a centrifugal column 2, and cracking the extracted sample under the action of a reagent solution 3;

3) with or without the addition of additive 9;

4) the collecting tube 1 with the centrifugal column 2 is placed in a centrifugal device for centrifugation, and liquid containing nucleic acid overcomes surface tension and drops into the collecting tube 1 from the centrifugal column 2 under the action of centrifugal force.

Example 2

As shown in fig. 4, the puncturing device comprises a puncturing rod 12 and a pressing head 15, the pressing head 15 is a rubber ball head or a plastic folding head, an opening is arranged in the middle of the test tube cover 7, and the storage tubes 8 are arranged in the inner cavity of the test tube cover 7 and connected into a whole through a connecting part; the pressing head 15 is positioned above the storage pipe 8, the upper part of the breaking rod 12 is fixedly connected with the inner upper wall of the pressing head 15, the breaking rod 12 downwards presses and breaks the bottom of the storage pipe 8 by pressing the pressing head 15, and meanwhile, the additive 9 is pushed into the centrifugal column 2 by the air pressure change of the pressing head 15. The other structure of this embodiment is as shown in embodiment 1.

Example 3

As shown in fig. 5, the puncturing device includes a puncturing rod 12, a rotating body 16 and a guide table 17, the middle of the test tube cap 7 is provided with an opening, the storage tube 8 is fixedly arranged below the opening, the rotating body 16 is positioned on the opening, the rotating body 16 is in rotating fit with the opening, the guide table 17 is arranged in the rotating body 16, the inner walls of the guide table 17 and the rotating body 16 are connected through threads, a reverse rotation guide convex strip 19 and a groove 18 are arranged between the lower part of the guide table 17 and the opening, the puncturing rod 12 is arranged below the guide table 17, the guide table 17 is driven to move downwards through the rotation of the rotating body 16, and the puncturing rod 12 pushes down the bottom of the storage tube 8. The guide table 17 may be provided with ventilation channels 20 on its inside or side walls, through which the additive falls by gravity into the spin column 2, in other configurations according to the embodiment shown in example 1.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention, including any reference to the above-mentioned embodiments. Various modifications to these embodiments will be readily apparent to those skilled in the art. The general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. The device is characterized by comprising a cover body and a puncturing device, wherein the cover body comprises a spiral pipe, a storage pipe (8) and a pressing stroke pipe, the spiral pipe is internally provided with threads, the pressing stroke pipe is positioned at the upper part of the spiral pipe, the storage pipe (8) is arranged in an inner cavity of the cover body and is connected with the cover body into a whole through a connecting part, and the bottom end of the storage pipe (8) is sealed; the breaking device comprises a breaking rod (12), a pressing head (13) and a sealing rubber piston (14), the breaking rod (12) and the pressing head (13) are connected with each other, the lower portion of the breaking rod (12) is located in the storage pipe (8), the pressing head (13) is arranged on the pressing stroke pipe, the sealing rubber piston (14) is fixedly arranged on the breaking rod (12), the outer wall of the sealing rubber piston (14) is attached to the inner wall of the storage pipe (8), the pressing head (13) is pressed downwards to enable the breaking rod (12) to break the bottom of the storage pipe (8), and pretreatment additives in the storage pipe (8) are pushed to flow out of the bottom of the storage pipe (8) through the sealing rubber piston (14).

2. The device for storing and adding the additive for the pretreatment of the biological sample as claimed in claim 1, wherein the upper part of the puncturing rod (12) is a guide rod (11), the guide rod (11) is matched with the inner wall of the storage tube (8), and the upper end of the guide rod (11) is fixedly connected with the pressing head (13).

3. The storage and addition device for additives in the pretreatment of biological samples is characterized in that the pressing head (13) is provided with a downward annular sleeve, the lower part of the annular sleeve is provided with a convex point (21) which protrudes out of the annular sleeve, the inner side of the upper part of the pressing stroke tube is provided with a notch (22) which is matched with the convex point (21), and the convex point (21) is clamped in the notch (22) to form positioning.

4. The device for storing and adding the additive for the pretreatment of the biological sample as claimed in claim 3, wherein a strip-shaped groove (23) is respectively formed on both sides of the convex point (21).

5. The device for storing and adding the pretreatment additive for the biological samples according to claim 1, wherein the pretreatment additive (9) is one or more of a lysis solution, a precipitation solution, a protein adsorption material and a protein precipitation material.

6. The device for storing and adding the pretreatment additive for the biological sample as claimed in claim 1, wherein the pretreatment additive (9) is a hemoglobin-adsorbing material.

7. The device for storing and adding the additive for the pretreatment of the biological sample as claimed in claim 1, wherein the sealing material (10) at the bottom of the storage tube (8) is aluminum foil or tin foil.

8. The device for storing and adding the additive for the pretreatment of the biological sample as claimed in claim 1, wherein the bottom of the piercing rod (12) is a pointed tip, and a liquid guide groove (24) is axially arranged above the pointed tip.

9. An integrated adding device for biological sample pretreatment reagents, which is characterized by comprising a collecting pipe (1), a centrifugal column (2) and a test tube cover (7), wherein the centrifugal column (2) is arranged in the collecting pipe (1), the test tube cover (7) is arranged at the upper end of the collecting pipe (1), and the test tube cover (7) adopts the storing and adding device according to any one of claims 1 to 8; after the bottom of the storage tube (8) is pierced, the pretreatment additive (9) in the storage tube (8) falls into the centrifugal column (2).