CN111286448A - A biological sample pretreatment additive storage and addition device - Google Patents

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

A biological sample pretreatment additive storage and addition device

technical field

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

Background technique

Real-time fluorescence quantitative PCR technology has been widely used in many fields such as molecular diagnosis of genetic diseases, clinical testing, animal and plant import and export quarantine, food safety monitoring, soil microbial detection and paternity testing. Since blood, food, soil and other samples contain a large number of inhibitory factors, such as hemoglobin, metheme, lactoferrin, humic acid, etc., they will have a significant inhibitory effect on conventional Taq DNA polymerase. Therefore, nucleic acids must be 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 the basis for downstream nucleic acid detection, and the extraction quality and integrity directly affect clinical research or diagnosis.

The general nucleic acid extraction process consists of three steps: sample lysis, binding and purification. The traditional extraction reagents have a relatively low acquisition rate and complicated steps, which have a certain impact on the extraction work. Especially when the sample amount is scarce, it is difficult to obtain qualified nucleic acids. In addition, the traditional method has many operation steps, high cost, large demand for samples, and is prone to cross-contamination, which is not conducive to the rapid extraction and purification of nucleic acid samples by customers.

In addition, in the most widely used real-time fluorescent PCR technology for nucleic acid detection at present, most of the reagents are not ready-to-use, resulting in the problems of complex operation, time-consuming and high cost in PCR detection. Errors and even failures in experiments. The operation of different personnel also causes experimental errors; one-step rapid nucleic acid extraction cannot be achieved; PCR detection reagents need to be manually configured on site, multiple steps need to be manually labeled, 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

In order to solve the above-mentioned technical problems, the purpose of the present application is to provide a biological sample pretreatment additive storage and addition device. The device adopts an additive that can be arranged in the storage tube, and can make the piercing rod pierce the storage tube by pressing the pressing head down. At the bottom of the storage tube, the pretreatment additive in the storage tube is pushed out from the bottom of the storage tube through the sealing rubber piston, which avoids the on-site configuration work and is convenient and quick to add.

In order to achieve the above-mentioned purpose, the application adopts the following technical solutions:

A biological sample pretreatment additive storage and addition device, the device includes a cover body and a breaking device, the cover body includes a spiral tube, a storage tube and a pressing stroke tube, the spiral tube is internally provided with threads, and the pressing stroke tube is located in the spiral tube. The upper part of the tube, the storage tube is arranged in the inner cavity of the cover body and is connected with the cover body through the connecting part, and 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 and the pressing head are connected to each other, the lower part of the piercing 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, and the outer wall of the sealing rubber piston is connected to the pressure stroke pipe. The inner wall of the storage tube is in contact with each other, and pressing down the pressing head can make the piercing rod penetrate the bottom of the storage tube, and push the pretreatment additive in the storage tube to flow out from the bottom of the storage tube through the sealing rubber piston.

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

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

As a further improvement, the two sides of the convex point are respectively provided with strip-shaped grooves, and the two-shaped grooves make the convex point located on an elastic sheet, which has good elasticity and is convenient for the pressing head to push down to break through the limit.

As a further improvement, the bottom of the piercing rod is a pointed tip, and a liquid guiding groove is axially arranged above the pointed tip, so that the liquid guiding groove will not cause the breaking rod and the damaged position of the bottom to stick closely, which will hinder the falling of the additive.

As a further improvement, the additive is one or more of a lysing solution, a purified solution, a protein adsorption material and a protein precipitation material. If the centrifuge tube is a preservation solution, the additive can be a lysing solution; if the centrifuge tube is a lysing solution, the additive can be one or more of purification solution, protein adsorption material and protein precipitation material, which will complete the lysis and purification in one step . Since blood, food, soil and other samples contain a large amount of hemoglobin, metheme, lactoferrin, humic acid, etc., it is further preferred that the additives are protein adsorption materials and protein precipitation materials; Said additive is hemoglobin adsorption material, which can avoid the influence of hemoglobin on fluorescent PCR.

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 also be thinned and sealed.

By adopting the above technical solution, the present application solves the technical problem that the additive in the storage tube may not completely fall into the spin column due to the difference in air pressure or surface tension after the bottom of the storage tube is broken, and the addition is convenient and quick.

Description of drawings

FIG. 1 is a schematic structural diagram of the application.

FIG. 2 is a schematic diagram of the explosion structure of the application.

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

FIG. 4 is a schematic structural diagram of a test tube cover in Example 2. FIG.

FIG. 5 is a schematic structural diagram of a test tube cover in Example 3. FIG.

Detailed ways

The specific embodiments of the present application will be described in detail below with reference to the accompanying drawings.

Example 1

As shown in FIG. 1 , a microbiological sample purification pretreatment device includes a collection tube 1 , a spin column 2 , a test tube cover 7 and a reagent solution 3 . The test tube cover 7 is connected with the collection tube 1 by threads, the centrifugal column 2 is arranged in the collection tube 1, the upper part of the centrifugal column 2 is provided with a convex ring 6, and the centrifugal column 2 is arranged on the upper part of the centrifugal column 2 through the convex ring 6. The upper end surface of the collection tube 1 and the lower part of the spin column 2 form a liquid collection cavity. The spin column 2 is arranged on the upper end face of the collection tube 1 through the convex ring 6 so that the spin column 2 is hung on the collection tube 1 , and the spin column 2 can be locked by setting the test tube cover 7 .

As shown in FIG. 1 , the test tube cover 7 is arranged on the upper end of the collection tube 1 to close the collection tube 1. The test tube cover 7 includes a cover body and a breaking device, and the cover body includes a spiral tube, a storage tube 8 and a pressing device. Stroke tube, the inside of the spiral tube is provided with threads, the pressing stroke tube is located on 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 through the connecting part, and the bottom end of the storage tube 8 is closed As shown in Figure 2, 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 protruding from the annular sleeve, and the upper inner side of the pressing stroke tube is provided with a convex point 21 to match The notch 22 is formed, the convex point 21 is clamped in the notch 22 to form a positioning, and the two sides of the convex point 21 are respectively provided with strip grooves 23 .

As shown in FIG. 1 , 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 guide rod 11, and the guide rod 11 is adapted to the inner wall of the storage tube 8. The upper end of the guide rod 11 is fixedly connected with the pressing head 13 . The lower part of the piercing rod 12 is located in the storage tube 8 , the lower part of the piercing rod 12 is located in the storage tube 8 , the bottom of the piercing rod 12 is a pointed tip, and a liquid guiding groove 24 is axially provided above the tip. 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 in contact with the inner wall of the storage pipe 8, and pressing the pressing head 13 can make the breaking rod 13 fit. 12 penetrates the bottom of the storage tube 8, and pushes the pretreatment additive in the storage tube 8 to flow out from the bottom of the storage tube 8 through the sealing rubber piston 14 and fall into the spin column 2.

As shown in FIG. 3 , the bottom of the spin 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 adopts a hydrophobic sieve plate, which is sintered from ultra-high molecular weight polyethylene (UHMW-PE), and the filtration precision of the hydrophobic sieve plate is 10-100 microns. The reagent solution 3 is arranged in the spin column 2 and is located above the surface tension layer 4, and 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 escape from the surface tension layer 4. pass. During the test, the whole set of device can be placed in a centrifuge, and when the spin column 2 is subjected to centrifugal force, the liquid will drop into the collection tube 1 from the spin column 2 against the surface tension. Alternatively, the test tube cover 7 can also be opened, and the liquid can be pushed into the collection tube 1 by setting the injection piston rod. Of course, under the condition of ensuring sufficient sealing between the test tube cover 7 , the spin column 2 and the collection tube 1 , the liquid can also be pushed into the collection tube 1 through the sealing rubber piston 14 at the upper end of the breaking device.

Taking nucleic acid extraction 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.

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

1) Add the reagent solution 3 to the spin column 2, or the spin column 2 itself presets the reagent solution 3;

2) Add the extract sample to the spin column 2, so that the extract sample is cleaved under the action of the reagent solution 3;

3) Add or not add additive 9;

4) Put the collection tube 1 with the spin column 2 into the centrifuge device for centrifugation. Through the action of centrifugal force, the nucleic acid-containing liquid overcomes the surface tension and drips into the collection tube 1 from the spin column 2.

Example 2

As shown in FIG. 4 , the piercing device includes a piercing rod 12 and a pressing head 15, the pressing head 15 adopts a rubber ball head or a plastic folding head, the test tube cover 7 is provided with an opening in the middle, and the storage tube 8 is arranged in the inner cavity of the test tube cover 7 and connected as a whole by the 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 with the inner upper wall of the pressing head 15, and by pressing the pressing head 15, The piercing rod 12 is pressed down to pierce the bottom of the storage tube 8 , and at the same time, the additive 9 is pushed into the spin column 2 by the change of the air pressure of the pressing head 15 . Other structures of this embodiment are as shown in Embodiment 1.

Example 3

As shown in FIG. 5 , the piercing device includes a piercing rod 12 , a rotating body 16 and a guide table 17 , an opening is provided in the middle of the test tube cover 7 , the storage tube 8 is fixedly arranged below the opening, and the rotating body 16 Located on the opening, the rotating body 16 and the opening are rotatably matched, the guide table 17 is arranged in the rotating body 16, the guide table 17 and the inner wall of the rotating body 16 are connected by threads, and a reverse guide is provided between the lower part of the guide table 17 and the opening The protruding strip 19 and the groove 18, the piercing rod 12 is arranged below the guide table 17, and the guide table 17 is driven to move downward by the rotating body 16. A ventilation channel 20 may be provided inside or on the side wall of the guide table 17 , and the additive falls into the spin column 2 by gravity. Other structures of this embodiment are as shown in Embodiment 1.

The above is a description of the embodiments of the present invention. The above description of the disclosed embodiments enables those skilled in the art to realize or use the present invention. Various modifications to these embodiments will be 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 present invention. Thus, the present invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novelties disclosed herein.

Claims (9)

1. A biological sample pretreatment additive storage and addition device, characterized in that the device comprises a cover body and a breaking device, the cover body comprises a spiral tube, a storage tube (8) and a pressing stroke tube, and the inside of the spiral tube is A thread is provided, the pressing stroke pipe is located on the upper part of the spiral pipe, the storage pipe (8) is arranged in the inner cavity of the cover body and is connected with the cover body through the connecting part, and the bottom end of the storage pipe (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) and the pressing head (13) are connected to each other, and the breaking rod (12) The lower part is located in 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 breaking rod (12), and the outer wall of the sealing rubber piston (14) is connected to the storage tube (14). The inner wall of the tube (8) is in contact with each other, and pressing the pressing head (13) down can make the breaking rod (12) penetrate the bottom of the storage tube (8), and push the front end of the storage tube (8) through the sealing rubber piston (14). The treatment additive flows out from the bottom of the storage tube (8). 2. A biological sample pretreatment additive storage and addition device according to claim 1, characterized in that the upper part of the breaking rod (12) is a guide rod (11), the guide rod (11) and the storage tube (8) ) inner wall, and the upper end of the guide rod (11) is fixedly connected with the pressing head (13). 3 . The biological sample pretreatment additive storage and addition device according to claim 1 , wherein the pressing head ( 13 ) is provided with a downward annular sleeve, and the lower part of the annular sleeve is provided with a protrusion protruding from the annular sleeve. 4 . A notch (22) adapted to the convex point (21) is provided on the upper inner side of the pressing stroke tube, and the convex point (21) is clamped in the notch (22) to form a positioning. 4. The biological sample pretreatment additive storage and addition device according to claim 3, characterized in that strip-shaped grooves (23) are respectively provided on both sides of the convex point (21). 5. A biological sample pretreatment additive storage and addition device according to claim 1, characterized in that, the pretreatment additive (9) is one of lysate, precipitation solution, protein adsorption material and protein precipitation material. one or more. 6. A biological sample pretreatment additive storage and addition device according to claim 1, characterized in that, the pretreatment additive (9) is a hemoglobin adsorption material. 7. The biological sample pretreatment additive storage and addition device according to claim 1, characterized in that, the bottom sealing material (10) of the storage tube (8) is aluminum foil or tin foil. 8. The biological sample pretreatment additive storage and addition device according to claim 1, wherein the bottom of the piercing rod (12) is a pointed tip, and a liquid guiding groove (24) is axially provided above the tip. ). 9. An integrated device for adding biological sample pretreatment reagents, characterized in that the device comprises a collection tube (1), a spin column (2) and a test tube cover (7), and the spin column (2) is provided in the collection tube (2) In the tube (1), the test tube cover (7) is arranged on the upper end of the collection tube (1), and the test tube cover (7) adopts the storage and addition device according to any one of claims 1-8. ; After the bottom of the storage tube (8) is broken, the pretreatment additive (9) in the storage tube (8) falls into the spin column (2).

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