Microfluidic stepwise multi-tube PCR reaction device

Abstract

The utility model relates to biological detection technical field, and disclose a kind of microfluidic step-by-step multi-tube PCR reaction device, comprising: for carrying out first round PCR reaction and second PCR reaction first test tube and second test tube, communication pipe is arranged between the first test tube and second test tube, and first test tube and second test tube are also communicated by communication pipe;The first test tube open end is also provided with a top cover, and flexible connecting strip is arranged between the top cover and the first test tube.The microfluidic step-by-step multi-tube PCR reaction device, the device is used for the nest PCR reaction needing step-by-step, simplifies the transfer process of sample between two-step PCR reaction, and in step-by-step PCR, the process that originally needs to open first PCR reaction tube first test tube and transfer product to second step PCR reaction tube communication pipe is completed in the device, without opening reaction tube, avoid the possible pollution brought by reaction tube opening.

Description

Technical Field

[0001] This utility model relates to the field of biological detection technology, specifically a microfluidic stepwise multitube PCR reaction device. Background Technology

[0002] PCR (polymerase chain reaction) is a technique used to amplify specific DNA fragments in vitro. It plays an important role in life science research and medical diagnosis. Nested PCR is a two-round PCR amplification technique. The two rounds of PCR are performed in steps. After the first round of PCR amplification, a small amount of product is taken as the template for the second round of PCR.

[0003] Nested PCR technology can greatly improve detection sensitivity due to its two-stage amplification process. It is also used in some diagnostic reagent products, such as the applicant's multiplex respiratory pathogen nucleic acid detection kit and multiplex meningitis / central nervous system infection pathogen detection kit. However, the process of transferring the product after the first round of PCR amplification into the second round of PCR reaction tube is prone to contamination and requires strict operation. To address this issue, we propose a microfluidic stepwise multitube PCR reaction device. Utility Model Content

[0004] To address the shortcomings of existing technologies, this invention provides a microfluidic stepwise multitube PCR reaction device, which solves the problems mentioned in the background.

[0005] This utility model provides the following technical solution: a microfluidic stepwise multi-tube PCR reaction device, comprising: a first test tube and a second test tube for performing a first round of PCR reaction and a second PCR reaction, wherein a connecting tube is provided between the first test tube and the second test tube, and the first test tube and the second test tube are also connected through the connecting tube; a top cover is provided at the open end of the first test tube, a flexible connecting strip is provided between the top cover and the first test tube, the two ends of the flexible connecting strip are respectively fixedly connected to the first test tube and the top cover, an annular sealing ring is fixedly installed on the inner side of the top cover, an annular groove is opened on the open end face of the first test tube, and a protrusion is provided on the outer surface of the top cover; a flow splitting mechanism is also provided on the top cover, the flow splitting mechanism includes a slide rod slidably connected to the top cover and an arc-shaped spring sheet fixedly installed on the top surface of the top cover, a pressure plate is fixedly installed at the upper end of the slide rod, and a squeezing head is fixedly installed at the lower end of the slide rod, and the upper end of the arc-shaped spring sheet is also fixedly connected to the pressure plate.

[0006] Preferably, the outer surface of the first test tube is engraved with scale lines, and the connection between the connecting tube and the first test tube is located above the scale lines.

[0007] Preferably, the shape of the annular groove is consistent with the shape of the annular sealing ring, and the annular sealing ring is made of rubber.

[0008] Preferably, the distance between the first test tube and the second test tube is equal to the distance between the two sockets in a standard PCR thermal cycler.

[0009] Preferably, the diversion mechanism further includes a retaining ring located at the connection between the arc-shaped spring and the top cover.

[0010] Preferably, both the first test tube and the second test tube are designed with a conical structure.

[0011] Preferably, the top cover is snapped into the first test tube, and after the top cover and the first test tube are snapped into place, the annular sealing ring is in close contact with the side of the annular groove.

[0012] Preferably, there are two arc-shaped spring pieces, which are symmetrically arranged on both sides of the slide bar.

[0013] Compared with the prior art, the present invention has the following beneficial effects:

[0014] 1. This microfluidic stepwise multi-tube PCR reaction device is used for nested PCR reactions that require stepwise execution. It simplifies the sample transfer process between two PCR reactions. In stepwise PCR, the process of transferring the product from the first PCR reaction tube to the connecting tube of the second PCR reaction tube, which originally required opening the first PCR reaction tube, is completed within this device without opening the reaction tube, thus avoiding potential contamination caused by opening the reaction tube.

[0015] 2. This microfluidic stepwise multitube PCR reaction device has the same external shape and size as ordinary PCR reaction tubes for the first test tube and connecting tube, which can be perfectly adapted to general PCR instruments on the market. No new instruments are needed when using this new device, and no additional costs are incurred. It can also be used for pathogen detection, genetic testing, tumor gene testing, etc. It is especially suitable for scenarios that require stepwise PCR for multiplex respiratory pathogen detection, multiplex meningitis pathogen detection, and point-of-care testing. The second step reaction can be carried out in multiple reaction tubes, and multiple connecting tubes are connected to the first test tube. It is especially suitable for multiplex respiratory pathogen detection and multiplex meningitis pathogen detection kits where the applicant needs multiple tubes in the second step. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the overall structure of this utility model;

[0017] Figure 2 This is a cross-sectional schematic diagram of the first test tube structure of this utility model;

[0018] Figure 3 This is a top view of the first state of the structure of this utility model;

[0019] Figure 4 This is a top view of the second state of the structure of this utility model;

[0020] Figure 5 This is a top view of the third state of the structure of this utility model.

[0021] In the diagram: 1. First test tube; 11. Scale line; 2. Connecting tube; 3. Second test tube; 4. Flexible connecting strip; 5. Top cover; 51. Protrusion; 52. Annular sealing ring; 53. Annular groove; 6. Diverting mechanism; 61. Extrusion head; 62. Slide rod; 63. Pressure plate; 64. Arc-shaped spring; 65. Snap ring. Detailed Implementation

[0022] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0023] Please see Figure 1-5 A microfluidic stepwise multi-tube PCR reaction device includes: a first test tube 1 and a second test tube 3 for performing a first round of PCR reaction and a second PCR reaction. A connecting tube 2 is provided between the first test tube 1 and the second test tube 3, and the first test tube 1 and the second test tube 3 are also connected through the connecting tube 2. The outer surface of the first test tube 1 is engraved with scale lines 11. The connection point between the connecting tube 2 and the first test tube 1 is located above the scale lines 11. The distance between the first test tube 1 and the second test tube 3 is equal to the distance between two sockets in a standard PCR thermal cycler. Both the first test tube 1 and the second test tube 3 are designed with a conical structure, so that the overall external shape and size are consistent with ordinary PCR reaction tubes, which can be perfectly adapted to general PCR instruments on the market. When using this new device, no new instruments are needed, and no additional costs are incurred.

[0024] The first test tube 1 is also provided with a top cover 5 at its open end. A flexible connecting strip 4 is provided between the top cover 5 and the first test tube 1. The two ends of the flexible connecting strip 4 are fixedly connected to the first test tube 1 and the top cover 5, respectively. An annular sealing ring 52 is fixedly installed on the inner side of the top cover 5. An annular groove 53 is opened on the open end face of the first test tube 1. A protrusion 51 is provided on the outer surface of the top cover 5. The shape of the annular groove 53 is consistent with the shape of the annular sealing ring 52. The annular sealing ring 52 is made of rubber. The top cover 5 is snapped into the first test tube 1. After the top cover 5 is snapped into the first test tube 1, the annular sealing ring 52 and the side of the annular groove 53 are in close contact. This can ensure the sealing of the overall structure after the top cover 5 is closed, and can prevent the sample inside the first test tube 1 from being contaminated.

[0025] The top cover 5 is also provided with a diversion mechanism 6. The diversion mechanism 6 includes a slide rod 62 that is slidably connected to the top cover 5 and an arc-shaped spring piece 64 that is fixedly installed on the top surface of the top cover 5. A pressure plate 63 is fixedly installed at the upper end of the slide rod 62, and an extrusion head 61 is fixedly installed at the lower end of the slide rod 62. The upper end of the arc-shaped spring piece 64 is also fixedly connected to the pressure plate 63. The diversion mechanism 6 also includes a retaining ring 65, which is located at the connection between the arc-shaped spring piece 64 and the top cover 5. There are two arc-shaped spring pieces 64 in total, which are symmetrically arranged on both sides of the slide rod 62. The arc-shaped spring pieces 64 can improve the overall performance of the mechanism and make its structure reusable.

[0026] Working principle: The first test tube 1 and the second test tube 3 are connected by a connecting tube 2, and the opening of the connecting tube 2 is located above the scale line 11 on the surface of the first test tube 1. Therefore, sufficient liquid can be injected into the first test tube 1 through the scale line 11. Then, the top cover 5 is locked onto the first test tube 1 to seal the first test tube 1 and the second test tube 3. Then, the pressure plate 63 is pushed down, and the squeezing head 61 slides down through the sliding rod 62, so that the squeezing head 61 slides into the liquid inside the first test tube 1. Thus, the liquid inside the first test tube 1 can flow into the second test tube 3 through the connecting tube 2 to transfer part of the product of the first step PCR reaction in the first test tube 1 to the second test tube 3 for the second step PCR reaction. After the pressure plate 63 is released, the pressure plate 63 drives the squeezing head 61 to slide back to its original position under the elastic force of the arc-shaped spring piece 64, which is fixedly connected to the pressure plate 63 and the top cover 5 at both ends.

[0027] Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A microfluidic stepwise multitube PCR reaction device, characterized in that, include: The first test tube (1) and the second test tube (3) are used for the first round of PCR reaction and the second PCR reaction. A connecting tube (2) is provided between the first test tube (1) and the second test tube (3), and the first test tube (1) and the second test tube (3) are also connected through the connecting tube (2). The first test tube (1) is also provided with a top cover (5) at the open end. A flexible connecting strip (4) is provided between the top cover (5) and the first test tube (1). The two ends of the flexible connecting strip (4) are fixedly connected to the first test tube (1) and the top cover (5) respectively. An annular sealing ring (52) is fixedly installed on the inner side of the top cover (5). An annular groove (53) is opened on the open end face of the first test tube (1). A protrusion (51) is provided on the outer surface of the top cover (5). The top cover (5) is also provided with a diversion mechanism (6). The diversion mechanism (6) includes a slide rod (62) that is slidably connected to the top cover (5) and an arc-shaped spring piece (64) that is fixedly installed on the top surface of the top cover (5). A pressure plate (63) is fixedly installed at the upper end of the slide rod (62), and an extrusion head (61) is fixedly installed at the lower end of the slide rod (62). The upper end of the arc-shaped spring piece (64) is also fixedly connected to the pressure plate (63).

2. The microfluidic stepwise multitube PCR reaction apparatus according to claim 1, characterized in that, The outer surface of the first test tube (1) is engraved with scale lines (11), and the connection between the connecting tube (2) and the first test tube (1) is located above the scale lines (11).

3. The microfluidic stepwise multitube PCR reaction apparatus according to claim 1, characterized in that, The shape of the annular groove (53) is consistent with the shape of the annular sealing ring (52), which is made of rubber.

4. The microfluidic stepwise multitube PCR reaction apparatus according to claim 1, characterized in that, The distance between the first test tube (1) and the second test tube (3) is equal to the distance between the two sockets in a standard PCR thermal cycler.

5. The microfluidic stepwise multitube PCR reaction apparatus according to claim 1, characterized in that, The diversion mechanism (6) also includes a retaining ring (65), which is located at the connection between the arc-shaped spring (64) and the top cover (5).

6. The microfluidic stepwise multitube PCR reaction apparatus according to claim 1, characterized in that, Both the first test tube (1) and the second test tube (3) are designed with a conical structure.

7. The microfluidic stepwise multitube PCR reaction apparatus according to claim 1, characterized in that, The top cover (5) is snapped into the first test tube (1), and after the top cover (5) and the first test tube (1) are snapped into place, the annular sealing ring (52) is in close contact with the side of the annular groove (53).

8. The microfluidic stepwise multitube PCR reaction apparatus according to claim 1, characterized in that, There are two arc-shaped spring pieces (64), which are symmetrically arranged on both sides of the slide bar (62).

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