Cover-opening-free double-system one-pot method detection device

Abstract

The utility model relates to the technical field of detection devices, and discloses an uncovering-free double-system one-pot method detection device which comprises a reaction mechanism, the reaction mechanism comprises a reaction tube and a cover body, the cover body covers the reaction tube to seal the reaction tube, the reaction tube and the cover body form a cavity, the cavity comprises a reaction area and a temporary storage area, and the temporary storage area is communicated with the reaction area. The reaction area is located below the temporary storage area, the reaction area is communicated with the temporary storage area, a liquid storage part is arranged in the temporary storage area, and liquid in the liquid storage part can be mixed with liquid in the reaction area (11) under the condition that the cover body is not opened. According to the utility model, the problems of potential risk of false positive caused by aerosol pollution and complexity of operation due to uncovering treatment of nucleic acid isothermal amplification products in the prior art can be solved.

Description

Technical Field

[0001] The utility model relates to the technical field of detection devices, in particular to a double-system one-pot detection device without opening the lid. Background Art

[0002] Since the 1990s, isothermal nucleic acid amplification technology (ITA) has developed rapidly. Currently reported isothermal nucleic acid amplification technologies include: transcription-mediated amplification, loop-mediated isothermal amplification, strand displacement amplification, helicase-dependent amplification, recombinase-polymerase amplification, etc.

[0003] ITA only requires a single incubation temperature, reducing the requirements for equipment. At the same time, it does not require repeated heating and cooling steps, reducing the reaction time. Most importantly, multiple molecular reactions can be carried out simultaneously instead of sequential operations in thermal cycling, which greatly increases the reaction efficiency. Given these advantages, isothermal nucleic acid amplification technology has been widely used in the detection of various pathogens.

[0004] However, the single ITA technology still has certain limitations, such as complex system primer design, only being able to amplify short fragments or circular DNA, complex detection of amplification products, low sensitivity or high false positives, etc. Therefore, ITA technology is usually combined with the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) - associated protein (Cas) system, enhancing the sensitivity, specificity, programmability, and visualization detection of the diagnostic technology.

[0005] However, the combination of ITA and the CRISPR / Cas system faces challenges in practical applications. The opening of the lid for ITA amplification products increases the potential risk of aerosol contamination leading to false positives and the complexity of operation. In addition, the interference between the RPA and CRISPR / Cas systems is not conducive to the storage, transportation, and application of the kit, etc. Summary of the Utility Model

[0006] In view of this, the purpose of the utility model is to provide a double-system one-pot detection device without opening the lid, solving the problems in the prior art that the opening of the lid for nucleic acid isothermal amplification products increases the potential risk of aerosol contamination leading to false positives and the complexity of operation.

[0007] The utility model solves the above technical problems through the following technical means:

[0008] A double-system one-pot detection device without opening the lid, comprising a reaction mechanism, the reaction mechanism includes a reaction tube and a lid, the lid covers the reaction tube to seal the reaction tube, the reaction tube and the lid form a cavity, the cavity includes a reaction area and a temporary storage area, the reaction area is located below the temporary storage area, the reaction area is communicated with the temporary storage area, a liquid storage member is arranged in the temporary storage area, and the liquid in the liquid storage member can be mixed with the liquid in the reaction area without opening the lid.

[0009] Further, the liquid storage member is a capillary tube, and the capillary tube is fixedly arranged in the temporary storage area.

[0010] Further, the liquid storage member is an adsorbent, and the adsorbent is fixedly arranged in the temporary storage area.

[0011] Further, a light condensing member is arranged on the lid, and the light condensing member is used for condensing the light emitted in the cavity.

[0012] Further, a connecting piece is arranged between the lid and the reaction tube, one end of the connecting piece is fixedly connected with the lid, and the other end is fixedly connected with the reaction tube.

[0013] Further, a plurality of the reaction mechanisms are provided, and the lids of the plurality of reaction mechanisms are installed on the same plate body.

[0014] Further, protrusions are arranged on both sides of the plate body close to the reaction tube at both ends.

[0015] Further, connecting rings are arranged on a plurality of the reaction tubes, and adjacent connecting rings are fixedly connected.

[0016] Further, a sealing ring is arranged in the lid, the sealing ring is inserted into the reaction tube, a convex ring is arranged at the end of the sealing ring, and an annular groove is formed on the inner wall of the reaction tube, and the convex ring is clamped in the annular groove.

[0017] Further, a marking area is arranged on the outer wall of the reaction tube.

[0018] The beneficial effects of the utility model:

[0019] By arranging the capillary tube, the utility model can store the two-system solutions at the bottom of the reaction tube and in the liquid storage member respectively. During detection, only the sample to be detected needs to be added. After the system solution at the bottom of the reaction tube reacts with the sample to be detected, the other system solution in the liquid storage member is then reacted with the sample to be detected. The mutual interference between the two-system solutions is effectively reduced; meanwhile, the operation is convenient and the lid does not need to be opened during the reaction process, avoiding the risk of aerosol contamination. Description of the Drawings

[0020] Figure 1 is the schematic plan view of a double - system one - pot detection device without opening the lid of the present utility model Figure 1 ;

[0021] Figure 2 is the schematic plan view of a double - system one - pot detection device without opening the lid of the present utility model Figure 2 ;

[0022] Figure 3 is the schematic sectional view of a double - system one - pot detection device without opening the lid of the present utility model;

[0023] Figure 4 is Figure 3 the enlarged schematic view of A in

[0024] Figure 5 is the schematic installation structure view of the condenser in a double - system one - pot detection device without opening the lid of the present utility model;

[0025] Figure 6 is the schematic connection structure view of multiple reaction mechanisms of a double - system one - pot detection device without opening the lid of the present utility model;

[0026] Figure 7 is the schematic structure view of the liquid storage member being set as an adsorbent and arranged at the inner top wall of the lid body;

[0027] Figure 8 is the schematic structure view of the liquid storage member being set as an adsorbent and arranged at the inner side wall of the lid body;

[0028] Among them,

[0029] 1. Reaction tube; 11. Reaction zone; 12. Temporary storage zone; 13. Annular groove; 14. Scale line; 15. Marking zone; 16. Connection ring;

[0030] 2. Lid body; 21. Sealing ring; 211. Convex ring;

[0031] 3. Capillary; 31. Flared mouth;

[0032] 4. Connection piece;

[0033] 5. Plate body; 51. Protrusion;

[0034] 6. Adsorbent;

[0035] 7. Condenser. Detailed implementation mode

[0036] The following describes the implementation manners of the present utility model through specific embodiments. Those skilled in the art can understand the advantages and effects of the present utility model from the content disclosed in this specification. It should be noted that the diagrams provided in the following embodiments are only for illustrative purposes, showing only schematic diagrams, not physical diagrams, and should not be construed as a limitation to the present utility model. In order to better illustrate the embodiments of the present utility model, some components in the diagrams will be omitted, enlarged or reduced, which do not represent the dimensions of the actual products; for those skilled in the art, it is understandable that some well-known structures and their descriptions in the diagrams may be omitted.

[0037] In the diagrams of the embodiments of the present utility model, the same or similar reference numerals correspond to the same or similar components. In the description of the present utility model, it should be understood that if there are terms such as "upper", "lower", "left", "right", "front", "rear", etc. indicating the orientation or position relationship, they are based on the orientation or position relationship shown in the diagrams. This is only for the convenience of describing the present utility model and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, the terms describing the position relationship in the diagrams are only for illustrative purposes and should not be construed as a limitation to the present utility model. For those of ordinary skill in the art, the specific meanings of the above terms can be understood according to specific circumstances.

[0038] As Figures 1 - 8 shown, a double-system one-pot detection device without opening the lid of the present utility model includes a reaction mechanism. The reaction mechanism includes a reaction tube 1 and a lid 2. The lid 2 covers the reaction tube 1 to seal the reaction tube 1. The lid 2 and the reaction tube 1 form a cavity. The cavity includes a reaction zone 11 and a temporary storage zone 12. The reaction zone 11 is located below the temporary storage zone 12. The reaction zone 11 is communicated with the temporary storage zone 12. A liquid storage member is provided in the temporary storage zone 12. The liquid in the liquid storage member can be mixed with the liquid in the reaction zone 11 without opening the lid 2.

[0039] As Figures 3 - 5 shown, in this embodiment, the liquid storage member is a capillary tube 3. The capillary tube 3 is fixedly arranged in the temporary storage zone 12. In this embodiment, the capillary tube 3 is fixedly installed on the lid 2. Specifically, the capillary tube 3 can be fixedly installed on the lid 2 by means of threaded connection, clamping, bonding, etc. In some other embodiments, the capillary tube 3 can also be fixedly installed on the side wall of the reaction tube 1 by means of threaded connection, clamping, bonding, etc. A clamping groove can also be opened on the inner wall of the reaction tube 1, and the capillary tube 3 can be clamped in the clamping groove. It should be noted that in this embodiment, the capillary tube 3 is a polytetrafluoroethylene capillary tube 3.

[0040] Before detecting the sample, prepare an isothermal amplification system (such as transcription-mediated amplification, loop-mediated isothermal amplification, strand displacement amplification, helicase-dependent amplification, recombinase polymerase amplification, etc.) solution, and add the RPA system solution to the reaction area 11 of reaction tube 1 (i.e., the bottom of the tube). Then, prepare a CRISPR / Cas (including: Cas12, Cas13, Cas14 and their similar gene editing enzymes) detection system solution, and add the CRISPR / Cas (including: Cas12, Cas13, Cas14 and their similar gene editing enzymes) detection system solution to capillary 3. During detection, add the sample to the reaction area 11 of reaction tube 1. After the sample reacts with the RPA system solution, centrifuge the detection device to mix the CRISPR / Cas (including: Cas12, Cas13, Cas14 and their similar gene editing enzymes) detection system solution in capillary 3 with the isothermal amplification system (such as transcription-mediated amplification, loop-mediated isothermal amplification, strand displacement amplification, helicase-dependent amplification, recombinase polymerase amplification, etc.) solution. After the reaction, observe the fluorescence signal with the naked eye under blue light, or place it in a fluorescence detector to monitor the fluorescence intensity generated by the reaction system in real time. The mutual interference between the two system solutions is effectively reduced; at the same time, the operation is convenient and there is no need to open the lid during the reaction process, avoiding the risk of aerosol contamination.

[0041] As Figure 5 shown, in this embodiment, a flared mouth 31 is provided at the end of the capillary to facilitate adding the CRISPR / Cas (including: Cas12, Cas13, Cas14 and their similar gene editing enzymes) detection system solution to capillary 3.

[0042] As Figures 7 - 8 shown, in some other embodiments, the liquid storage member is an adsorbent 6, and the adsorbent 6 can be set as a sponge, cotton cloth, cotton strip, etc. The adsorbent 6 is fixedly arranged in the temporary storage area 12. Before detecting the sample, prepare an isothermal amplification system solution, and add the isothermal amplification system solution to the reaction area 11 of reaction tube 1 (i.e., the bottom of the tube). Then, prepare a CRISPR / Cas detection system solution, and add the CRISPR / Cas detection system solution to the adsorbent 6. During detection, add the sample to the reaction area 11 of reaction tube 1. After the sample reacts with the isothermal amplification system solution, invert the detection device to mix the isothermal amplification system solution in the reaction area 11 with the CRISPR / Cas detection system solution adsorbed in the adsorbent 6. After the reaction, observe the fluorescence signal with the naked eye under blue light, or place it in a fluorescence detector to monitor the fluorescence intensity generated by the reaction system in real time.

[0043] In some other embodiments, the adsorbent 6 can be arranged on the inner top wall of the cover body 2 by clamping, bonding, etc. (such as Figure 7As shown, the adsorbent 6 can also be arranged on the inner side wall of the cover body 2 by clamping, bonding, etc. (such as Figure 8 shown), the adsorbent 6 can also be arranged on the inner wall of the upper end of the reaction tube 1 by clamping, bonding, etc., which can meet the requirement that when the detection device is inverted, the isothermal amplification system solution in the reaction zone 11 is mixed with the CRISPR / Cas detection system solution adsorbed in the adsorbent 6.

[0044] Such as Figure 5 , 7 and 8 shown, in this embodiment, a light condensing member 7 is arranged on the cover body 2, and the light condensing member 7 is used to converge the light emitted in the cavity, so as to facilitate observing the fluorescence intensity generated in the cavity from the top of the detection device. In some other embodiments, the light condensing member 7 can be set as a convex lens and fixedly connected to the cover body 2 through a notch opened on the cover body 2. In this embodiment, the cover body 2 is made of a light-transmitting material, and the light condensing member 7 is set as two semi-convex lenses, and the two semi-convex lenses are respectively fixedly arranged on the inner and outer sides of the top of the cover body 2. It should be noted that the end faces of the two semi-convex lenses can be set as annular or circular according to the type of the liquid storage member.

[0045] Such as Figure 3 and Figure 4 shown, in this embodiment, a sealing ring 21 is arranged in the cover body 2, the sealing ring 21 is inserted into the reaction tube 1, a convex ring 211 is arranged at the end of the sealing ring 21, and an annular groove 13 is opened on the inner wall of the reaction tube 1, and the convex ring 211 is clamped in the annular groove 13. The sealing between the cover body 2 and the reaction tube 1 is realized through the sealing ring 21, and the clamping between the cover body 2 and the reaction tube 1 is realized through the convex ring 211. In some other embodiments, the sealing ring 21 can also be set as a cone to realize sealing and clamping. In this embodiment, the convex ring 211 is used in cooperation with the annular groove 13, which can ensure the clamping effect and avoid the situation that the cover body 2 and the reaction tube 1 are automatically separated.

[0046] Such as Figure 1 shown, in this embodiment, a scale line 14 is arranged on the outer wall of the reaction tube 1, which is convenient for the tester to observe the amount of the solution in the reaction tube 1. A marking area 15 is arranged on the outer wall of the reaction tube 1. The marking area 15 is a frosted area, which is convenient for the tester to mark the reaction tube 1.

[0047] Such as Figure 1 and Figure 2 shown, in this embodiment, a connecting piece 4 is arranged between the cover body 2 and the reaction tube 1. The connecting piece 4 is made of a ductile material. One end of the connecting piece 4 is fixedly connected to the cover body 2, and the other end is fixedly connected to the reaction tube 1. The cover body 2 is connected to the reaction tube 1 through the connecting piece 4 to prevent the cover body 2 from being lost.

[0048] Such as Figure 6As shown, in some other embodiments, a plurality of reaction mechanisms are provided. The covers 2 of the plurality of reaction mechanisms are installed on the same plate body 5, which can also prevent the loss of the covers 2. Protrusions 51 are provided on both ends of the plate body 5 near the reaction tubes 1. By pressing the protrusions 51, it is more convenient to disassemble the plate body 5 and the cover 2. At the same time, connection rings 16 are provided on all the reaction tubes 1, and the adjacent connection rings 16 are fixedly connected. Multiple reaction tubes 1 can be moved simultaneously without operating each reaction tube 1 one by one, improving convenience.

[0049] The working principle of the present utility model is as follows:

[0050] First, prepare an isothermal amplification system solution and add the isothermal amplification system solution to the reaction zone 11 of the reaction tube 1.

[0051] Then, prepare a CRISPR / Cas detection system solution and add the CRISPR / Cas detection system solution to the capillary 3.

[0052] Subsequently, add the sample to be detected into the reaction tube 1 to make the sample to be detected react with the RPA system solution.

[0053] After the reaction between the sample to be detected and the isothermal amplification system solution is completed, centrifuge the detection device to mix the CRISPR / Cas detection system solution in the capillary 3 with the RPA system solution. After the reaction is completed, observe the fluorescence signal.

[0054] The above embodiments are only used to illustrate the technical solutions of the present utility model and not to limit them. Although the present utility model has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present utility model can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present utility model, and they should all be covered within the scope of the claims of the present utility model. The technologies, shapes, and structures not described in detail in the present utility model are all well-known technologies.

Claims

1. A double-system one-pot detection device without opening the lid, characterized in that: It includes a reaction mechanism, the reaction mechanism includes a reaction tube (1) and a cover body (2), the cover body (2) covers the reaction tube (1) to seal the reaction tube (1), the reaction tube (1) and the cover body (2) form a cavity, the cavity includes a reaction zone (11) and a temporary storage zone (12), the reaction zone (11) is located below the temporary storage zone (12), the reaction zone (11) communicates with the temporary storage zone (12), a liquid storage member is arranged in the temporary storage zone (12), and the liquid in the liquid storage member can be mixed with the liquid in the reaction zone (11) without opening the cover body (2).

2. The dual-system one-pot detection device without opening the lid according to claim 1, characterized in that: The liquid storage member is a capillary tube (3), and the capillary tube (3) is fixedly arranged in the temporary storage zone (12).

3. The double-system one-pot detection device without opening the lid according to claim 1, wherein: The liquid storage member is an adsorbent (6), and the adsorbent is fixedly arranged in the temporary storage zone (12).

4. The dual-system one-pot detection device without opening the lid according to claim 1, characterized in that: A light condensing member (7) is arranged on the cover body, and the light condensing member (7) is used for condensing the light emitted in the cavity.

5. The dual-system one-pot detection device without opening the lid according to claim 1, characterized in that: A connecting piece (4) is arranged between the cover body (2) and the reaction tube (1), one end of the connecting piece (4) is fixedly connected with the cover body (2), and the other end is fixedly connected with the reaction tube (1).

6. The double-system one-pot detection device without opening the lid according to claim 1, characterized in that: A plurality of the reaction mechanisms are arranged, and the cover bodies (2) of the plurality of reaction mechanisms are installed on the same plate body (5).

7. The double-system one-pot detection device without opening the lid according to claim 6, wherein: Protrusions (51) are arranged on both sides of the two ends of the plate body (5) close to the reaction tube (1).

8. The double-system one-pot detection device without opening the lid according to claim 7, characterized in that: Connecting rings (16) are arranged on a plurality of the reaction tubes (1), and adjacent connecting rings (16) are fixedly connected to each other.

9. A non-opening-required dual-system one-pot detection device according to any one of claims 1-8, characterized in that: A sealing ring (21) is arranged in the cover body (2), the sealing ring (21) is inserted into the reaction tube (1), a convex ring (211) is arranged at the end of the sealing ring (21), and an annular groove (13) is formed in the inner wall of the reaction tube (1), and the convex ring (211) is clamped in the annular groove (13).

10. A double-system one-pot detection device without opening the lid according to claim 1, characterized in that: A marking area (15) is arranged on the outer wall of the reaction tube (1).

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