On-site visual detection device based on recombinase polymerase amplification

Abstract

The present invention relates to the technical field of nucleic acid amplification product detection, and in particular to an on-site visualization detection device based on RPA, comprising a reaction tube, a tube support, a heating plate and a syringe; a tube cover is provided on the top of the reaction tube, the reaction tube is provided on the tube support, a heating area is formed between the lower part of the reaction tube and the tube support, and a heating plate is provided in the heating area; the syringe comprises a puncture needle and a barrel body, the puncture needle can pierce the tube cover, a transparent visual window is provided on the side wall of the barrel body, a cavity is provided in the visual window, and a test paper is provided in the cavity; a button movable in the horizontal direction is provided on the visual window, the outer end of the button is located outside the visual window, the inner end of the button is located in the cavity and corresponds to the position of the test paper, and the inner end of the button can pierce the side wall of the syringe. The present invention is easy to operate and easy to observe experimental results, which meets the needs of on-site rapid detection.

Description

Technical Field

[0001] The present invention relates to the technical field of nucleic acid amplification product detection, and in particular to an on-site visual detection device based on RPA. Background Art

[0002] In recent years, with the development of molecular biology technology, more and more nucleic acid amplification technologies have been applied to the detection of pathogens. Recombinant enzyme polymerase amplification (RPA) technology is a new type of nucleic acid reaction technology that can replace PCR. RPA uses the T4 phage nucleic acid replication mechanism as the main reaction principle, and uses recombinase, single-stranded binding protein and DNA polymerase to replace the thermal cycle denaturation process of traditional PCR, realizing rapid nucleic acid amplification under constant temperature conditions (25℃-42℃), and the amplified product can be obtained in 20 minutes. It has good operability and can complete nucleic acid detection under non-laboratory conditions. It is considered to be the closest to the so-called "isothermal amplification" and has broad application prospects.

[0003] Although the constant temperature amplification technology ensures the rapid amplification of nucleic acids, it cannot be well connected with the later result judgment. At present, fluorescent dyes are often added after amplification, which increases the operation steps and the risk of aerosol contamination; another method is to use fluorescent instruments to detect the amplification signal. These methods cannot fully meet the needs of on-site rapid detection. Summary of the invention

[0004] To solve the above problems, the purpose of the present invention is to provide an on-site visual detection device based on RPA. After the reaction is completed, the reaction reagent in the reaction tube is extracted through the syringe, and the test paper on the side wall of the syringe is pressed into the syringe by pressing the button. By observing the changes in the test paper, the result can be judged, which is conducive to rapid on-site detection.

[0005] For the purpose of the present invention, the following technical solutions are adopted to implement:

[0006] A field visualization detection device based on RPA, the device comprises a reaction tube, a tube support, a heating plate and a syringe; a tube cover is arranged on the top of the reaction tube, the reaction tube is arranged on the tube support, a heating area is formed between the lower part of the reaction tube and the tube support, and the heating plate is arranged in the heating area; the syringe comprises a puncture needle and a tube body, the puncture needle can pierce the tube cover, a transparent visual window is arranged on the side wall of the tube body, a cavity is arranged in the visual window, and a test paper is arranged in the cavity; a button movable in the transverse direction is arranged on the visual window, the outer end of the button is located outside the visual window, the inner end of the button is located in the cavity and corresponds to the position of the test paper, and the inner end of the button can pierce the side wall of the tube body.

[0007] Preferably, the button is near the bottom of the cartridge body.

[0008] Preferably, the inner end of the button is needle-point shaped.

[0009] Preferably, a wing plate is provided at the outer end of the button, and the diameter of the wing plate is larger than the button.

[0010] Preferably, the material of the tube cover is rubber.

[0011] Preferably, the reaction tube is an eight-tube junction, and the tube caps are interconnected.

[0012] Preferably, the tube support includes a horizontal support plate and a vertical plate. The horizontal support plate is provided with a mounting hole extending vertically therethrough, and the reaction tube is arranged in the mounting hole. Vertical plates are arranged on the front and rear sides of the horizontal support plate, and the lower end of the vertical plates is located below the bottom of the reaction tube. A U-shaped heating area is formed between the reaction tube and the front and rear vertical plates.

[0013] Preferably, the pipe support is a cuboid with side openings.

[0014] Preferably, a downwardly extending partition is provided at the bottom of the horizontal support plate, the partition is located at the front and rear sides of the mounting hole, and the upper end of the heating plate is located on the outside of the two partitions.

[0015] Preferably, the viewing window extends along the length direction of the outer wall of the barrel body.

[0016] In summary, the advantages of the present invention are: after the reaction is completed, the syringe penetrates the tube cover and is inserted into the reaction tube to directly extract the reaction reagent, thereby preventing aerosol contamination. A transparent visual window is provided on the side wall of the syringe, and a test paper is provided in the visual window. By pressing a button, the test paper can be pushed into the interior of the syringe, and the result can be judged by observing the reaction of the test paper. The operation is convenient and the experimental results can be easily observed, which meets the needs of rapid on-site detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Schematic diagram of the structure of the reaction tube and the tube support.

[0018] Figure 2 A side view of the reaction tube and tube holder.

[0019] Figure 3 It is a schematic diagram of the structure of the syringe.

[0020] Figure 4 for Figure 3 A partial enlarged view of point A in the middle. DETAILED DESCRIPTION

[0021] like Figures 1 to 4As shown, an on-site visualization detection device based on RPA includes a reaction tube 1, a tube holder 2, a heating plate 3 and a syringe 4. The reaction tube 1 is filled with a reaction reagent, and a tube cover 11 is arranged on the top of the reaction tube 1, and the tube cover 11 seals the reaction reagent in the reaction tube 1. Preferably, the material of the tube cover 11 is rubber. The reaction tube 1 is an eight-tube, and each tube cover 11 is connected to each other. The reaction tube 1 is arranged on the tube holder 2, and a heating area is formed between the lower part of the reaction tube 1 and the tube holder 2. A heating plate 3 is arranged in the heating area, and the heating plate 3 is used to heat the reaction tube 1 so that the reagent in the reaction tube 1 undergoes an amplification reaction.

[0022] like Figure 1 and Figure 2 As shown, the tube support 2 includes a horizontal support plate 21 and a vertical plate 22. The horizontal support plate 21 is provided with a mounting hole that runs through from top to bottom, and the reaction tube 1 is arranged in the mounting hole 21. Vertical plates 22 are arranged on the front and rear sides of the horizontal support plate 21, and the lower ends of the vertical plates 22 are located below the bottom of the reaction tube 1. A U-shaped heating area is formed between the reaction tube 1 and the front and rear vertical plates 22, which is conducive to increasing the contact area between the heating plate 3 and the reaction tube 1 and improving the heating effect. Preferably, the tube support 2 is a rectangular parallelepiped with a side opening, and the heating plate 3 is placed into the heating area from the opening.

[0023] like Figure 2 As shown, a downwardly extending partition 211 is provided at the bottom of the horizontal support plate 21 , and the partition 211 is located at the front and rear sides of the mounting hole, and the upper end of the heating plate 3 is located outside the two partitions 211 .

[0024] like Figure 3 and Figure 4 As shown, the syringe 4 includes a puncture needle 41 and a barrel body 42. The puncture needle 41 can pierce the tube cover 11. A transparent visual window 5 is provided on the side wall of the barrel body 42. A cavity is provided in the visual window 5. A test paper is provided in the cavity. The test paper is in a long strip shape. A button 6 that moves in the horizontal direction is provided on the visual window 5. The outer end of the button 6 is located outside the visual window 5. The inner end of the button 6 is located in the cavity and corresponds to the position of the test paper. When in use, the outer end of the button 6 is pressed so that the inner end of the button 6 is pressed on the test paper. After piercing the barrel body 42, the test paper is pushed into the interior of the barrel body 42.

[0025] Preferably, the button 6 is close to the bottom of the barrel body 42 to ensure that the test paper can contact the detection reagent in the syringe 4. The inner end of the button 6 is in the shape of a needle tip, which is convenient for piercing the side wall of the barrel body 42 so that the test paper enters the barrel body 42. The outer end of the button 6 is provided with a wing plate 7, and the diameter of the wing plate 7 is larger than the button 6 to prevent the entire button 6 from being pressed into the visual window 5. The visual window 5 extends along the length direction of the outer wall of the barrel body 42, which is convenient for observing the reaction of the test paper.

[0026] When the device is used: after the system configuration and subpackaging into the eight-tube strip, the eight-tube strip is placed in the tube holder 2, and then a heating plate 3 (warming patch) is placed in the tube holder 2 to form a temperature zone around the eight-tube strip, and heating is performed to promote the amplification reaction. After 20 minutes of reaction, the syringe 4 with the test strip is inserted into the rubber area on the cover of the eight-tube strip, the reaction reagent is extracted and appropriate pure water is extracted for dilution, and finally the test strip is pushed into the syringe through the button 6 at the bottom of the side of the syringe 4 to contact the diluted reagent, triggering the test strip reaction, and the final result can be observed through the visual window on the side. If two red lines appear, it is positive, and one red line is negative.

[0027] The device has the following advantages: after the reaction is completed, the syringe 4 penetrates the tube cover 11 and is inserted into the reaction tube 1 to directly extract the reaction reagent, thereby preventing aerosol contamination. A transparent visual window 5 is provided on the side wall of the syringe 4, and a test paper is provided in the visual window 5. By pressing the button 6, the test paper can be pushed into the syringe 4, and the result can be judged by observing the reaction of the test paper. The operation is convenient and the experimental results can be easily observed, which meets the needs of rapid on-site detection.

Claims

1. An on-site visual detection device based on recombinase polymerase amplification, characterized in that: The device comprises a reaction tube (1), a tube support (2), a heating plate (3) and a syringe (4); a tube cover (11) is arranged on the top of the reaction tube (1); the reaction tube (1) is arranged on the tube support (2); a heating area is formed between the lower part of the reaction tube (1) and the tube support (2); the heating plate (3) is arranged in the heating area; the syringe (4) comprises a puncture needle (41) and a syringe body (42); the puncture needle (41) can puncture the tube cover (11); a transparent visual window (5) is arranged on the side wall of the syringe body (42); a transparent visual window (5) is arranged in the visual window (5); A cavity is provided, and a test paper is provided in the cavity; a button (6) movable in the transverse direction is provided on the visual window (5), the outer end of the button (6) is located outside the visual window (5), the inner end of the button (6) is located in the cavity and corresponds to the position of the test paper, and the inner end of the button (6) can pierce the side wall of the barrel body (42); the button (6) is close to the bottom of the barrel body (42); when in use, the outer end of the button is pressed so that the inner end of the button is pressed on the test paper, and after the button pierces the barrel body, the test paper is pushed into the interior of the barrel body, and the test paper contacts the detection reagent in the barrel body.

2. The on-site visual detection device based on recombinase polymerase amplification according to claim 1, characterized in that: The inner end of the button (6) is in the shape of a needle tip.

3. The on-site visual detection device based on recombinase polymerase amplification according to claim 1, characterized in that: A wing plate (7) is provided at the outer end of the button (6), and the diameter of the wing plate (7) is larger than the diameter of the button (6).

4. The on-site visual detection device based on recombinase polymerase amplification according to claim 1, characterized in that: The material of the pipe cover (11) is rubber.

5. The on-site visual detection device based on recombinase polymerase amplification according to claim 1, characterized in that: The reaction tube (1) is an eight-tube connection, and the tube caps (11) are connected to each other.

6. The on-site visual detection device based on recombinase polymerase amplification according to claim 1, characterized in that: The tube support (2) comprises a horizontal support plate (21) and a vertical plate (22); the horizontal support plate (21) is provided with a mounting hole penetrating vertically, and the reaction tube (1) is arranged in the mounting hole (21); the vertical plates (22) are arranged on the front and rear sides of the horizontal support plate (21), and the lower ends of the vertical plates (22) are located below the bottom of the reaction tube (1); and a U-shaped heating area is formed between the reaction tube (1) and the front and rear vertical plates (22).

7. The on-site visual detection device based on recombinase polymerase amplification according to claim 6, characterized in that: The pipe support (2) is a rectangular parallelepiped with an opening on the side.

8. The on-site visual detection device based on recombinase polymerase amplification according to claim 6, characterized in that: A downwardly extending partition (211) is provided at the bottom of the horizontal support plate (21), the partition (211) is located at the front and rear sides of the mounting hole, and the upper end of the heating plate (3) is located outside the two partitions (211).

9. The on-site visual detection device based on recombinase polymerase amplification according to claim 1, characterized in that: The viewing window (5) extends along the length direction of the outer wall of the cylinder body (42).

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