Constant-temperature nucleic acid amplification detection quantitative transfer consumables and detection equipment

Abstract

The utility model relates to a constant temperature nucleic acid amplification detection quantitative transfer consumptive material, detection equipment, wherein constant temperature nucleic acid amplification detection quantitative transfer consumptive material includes: detection unit, at least is equipped with the detection cup of first reagent and can receive sample preloading, pipetting unit has the channel of second reagent prestorage, and at least has a first vesicle with the intercommunication of channel, the pipetting unit with detection unit is connected through the channel with detection cup, forms the closed communication chamber, and when the first vesicle is squeezed, second reagent can enter the detection cup through the channel. The quantitative transfer consumptive material of the application can accurately quantitatively transfer liquid, can block aerosol pollution when detecting all the time zero opening, significantly reduces false positive, liquid transfer volume is fixed, error is small, and quantitative is accurate, and can be compatible with the existing standard tube type reaction system, and disposable use avoids cross contamination.

Description

Technical Field

[0001] This application belongs to the field of medical molecular diagnostic consumables technology, and in particular relates to a detection consumable and detection equipment that can complete nucleic acid amplification reaction under constant temperature conditions and can quantitatively transfer reagents in a sealed manner. Background Technology

[0002] Isothermal amplification technologies, such as loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and nicking enzyme amplification reaction (NEAR), have been widely used in primary healthcare institutions, port quarantine, and point-of-care testing (POCT) due to their advantages such as not requiring a thermal cycler, short detection time, and high sensitivity. However, current isothermal amplification testing consumables have many shortcomings in their design and application.

[0003] First, the numerous steps involved in opening the container and adding reagents easily lead to aerosol contamination, resulting in false positives and affecting detection accuracy. Second, manual pipetting has significant errors, especially with micro-volume reagents (10-100µL), making precise quantification difficult and causing inconsistencies in the reaction system. Third, lyophilized reagents require on-site reconstitution, and the reconstitution volume is difficult to control, further affecting the reproducibility and stability of the reaction. Fourth, poor optical path design in the reaction vessel results in low sensitivity for colorimetric or fluorescence signal acquisition, making it difficult to detect low-copy nucleic acids. Fifth, multi-component reagents need to be added stepwise, which is complex and hinders the achievement of a single-stage closed reaction, increasing the risk of contamination and operational difficulty.

[0004] While existing technologies have attempted to address these issues using microfluidic chips and sealing membranes, microfluidic chips are costly to manufacture and difficult to adapt to standard 50-100 mL reaction systems. Products utilizing siphon-based quantitative structures also exist, but these are mostly open designs and still fail to solve the problems of aerosol contamination and the need for stepwise reagent addition. Therefore, there is an urgent need in this field for a low-cost, disposable, precisely quantitative, and fully sealed isothermal amplification detection consumable to overcome the shortcomings of existing technologies. Summary of the Invention

[0005] This application provides a quantitative transfer consumable for isothermal nucleic acid amplification detection, solving the technical problem of how to provide a simple, low-cost, disposable isothermal nucleic acid amplification detection consumable that can be used immediately after opening the cap, with pre-filled lyophilized reagents.

[0006] To solve at least one of the above-mentioned technical problems, the technical solution adopted in this application is:

[0007] A quantitative transfer consumable for isothermal nucleic acid amplification detection includes:

[0008] The detection unit is equipped with at least one detection cup that can be pre-loaded with the first reagent and can receive the sample;

[0009] The pipetting unit has a channel for pre-storing a second reagent and at least one first vesicle communicating with the channel;

[0010] The pipetting unit and the detection unit are connected to the detection cup through the channel to form a closed and connected chamber; and when the first vesicle is squeezed, the second reagent can enter the detection cup through the channel.

[0011] Furthermore, the detection cup has a conical structure, comprising a lower first cone, a middle second frustum, and an upper cylinder; the generatrix angle of the first cone is greater than the generatrix angle of the second frustum.

[0012] Furthermore, the volume of the detection cup is greater than the volume of the first reagent; and when the first vesicle is squeezed, the channel is inserted into the first cone, so that the second reagent is squeezed into the first cone.

[0013] Furthermore, the test cup is also provided with a sealing cap that seals with it, and the sealing cap is detachably connected to the test cup.

[0014] Furthermore, the channel is integrally injection molded with the sealing cap, or it is assembled separately and then fixed to the lower surface of the sealing cap.

[0015] Furthermore, the pipetting unit also includes a second vesicle, and the first vesicle and the second vesicle are interconnected.

[0016] Furthermore, the first vesicle and the second vesicle are both located on the same side of the channel; and the first vesicle, the second vesicle, and the channel are integrally connected.

[0017] Furthermore, one end of the second vesicle is connected to the channel, and the other end is connected to the first vesicle.

[0018] Furthermore, the second vesicle is located at the end of the channel away from the detection cup, and the first vesicle is located above the second vesicle.

[0019] An isothermal nucleic acid amplification and detection device, adapted to the quantitative transfer consumables described above.

[0020] This application discloses a isothermal nucleic acid amplification and detection quantitative transfer consumable and detection device. The device eliminates the need to open the cap during the entire detection process, effectively blocking aerosol contamination and significantly reducing false positives. The channel has a fixed volume, allowing for precise quantitative liquid transfer. The detection cup can be pre-filled with lyophilized reagents, supporting room temperature storage and transportation; it is ready to use immediately after opening, eliminating the need for cold chain and precise liquid transfer steps. The detection cup also features a conical bottom and high light transmittance design, enhancing fluorescence signal by >20%, making it suitable for low-copy nucleic acid detection. It is compatible with existing standard tubular reaction systems, and its single-use design avoids cross-contamination. The overall structure is simple, allowing for integrated injection molding or modular assembly; the process is mature and suitable for large-scale production. Attached Figure Description

[0021] Figure 1 This is an exploded view of the consumables in this application;

[0022] Figure 2 This is an example diagram showing the consumables in this application combined together;

[0023] Figure 3 This is a cross-sectional view of the consumables in this application assembled together;

[0024] Figure 4 This is an example diagram of the test cup and sealing cap combined in this application;

[0025] Figure 5 This is a cross-sectional view of the test cup and sealing cap combined in this application;

[0026] Figure 6 This is an example diagram showing the integral molding of the pipetting unit and the sealing cap in this application;

[0027] Figure 7 This is a schematic diagram showing the usage state before liquid transfer in this application;

[0028] Figure 8 This is a schematic diagram showing the usage state after liquid transfer, as described in this application.

[0029] In the picture:

[0030] 10. Detection Unit 11. Testing cup 111. First cone 112. Second frustum 113. Cylinder 12. Sealing cap 20. Pipetting Unit 21. Passage 22. First vesicle 23. Second vesicle Detailed Implementation

[0031] The present application will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0032] This embodiment proposes a quantitative transfer consumable for isothermal nucleic acid amplification detection, such as... Figure 1-3As shown, the device includes a detection unit 10 and a pipetting unit 20. The detection unit 10 is equipped with at least one detection cup 11 for pre-filling a first reagent and for receiving samples. The pipetting unit 20 has a channel 21 for pre-storing a second reagent and at least one first vesicle 22 communicating with the channel 21. The pipetting unit 20 and the detection unit 10 are connected to the detection cup 11 via the channel 21, forming a closed, interconnected chamber. When the first vesicle 22 is squeezed, the second reagent can enter the detection cup 11 through the channel 21. After the detection unit 10 and the pipetting unit 20 are connected, a closed chamber is formed. By squeezing the first vesicle 22, the second reagent can be quantitatively and tightly transferred to the detection cup 11, achieving zero-opening operation, thereby avoiding aerosol contamination and improving detection accuracy.

[0033] like Figure 4-5 As shown, the detection cup 11 has a conical structure, comprising a lower first cone 111, a middle second frustum 112, and an upper cylinder 113. The channel 21 is inserted into the first cone 111, allowing the second reagent to be squeezed into it. The detection cup 11 adopts a three-section conical structure, with the outlet of the pipetting channel 21 extending into the interior of the lower cone 111. This design ensures that the second reagent is directly squeezed into the reaction area, i.e., accurately delivered into the lower cone 111; moreover, the conical structure facilitates liquid concentration, prevents reagent adhesion to the walls, and reduces residue; it also facilitates rapid and thorough mixing of the reagent with the pre-filled lyophilized reagent and sample at the bottom of the cup.

[0034] The generatrix angle θ of the first cone 111 is greater than the generatrix angle β of the second frustum 112, thus forming a steeper cone bottom. This steep cone bottom structure is mainly used for pre-loading and carrying lyophilized reagents. This design ensures that the reagents can be stably concentrated at the bottom of the cup, making it easy to use on-site without complicated reconstitution operations, thus improving the convenience and reliability of the test.

[0035] The lower end of the first cone 111, which forms the lower section, is a closed arc-shaped structure with a generatrix angle θ ranging from 20° to 30°, including but not limited to 20°, 21°, 22°, 23°, 24°, 25°, 26°, 27°, 28°, 29°, and 30°. It is primarily used for pre-filling the first reagent, which is a lyophilized reagent, such as lyophilized microspheres, ready for immediate use upon opening. The first cone 111 utilizes its steep cone angle to form a concentrated reaction zone, mainly used for pre-filling and containing the lyophilized reagent, and serving as the outlet for the pipetting channel, ensuring that the reagent is accurately added to the core area.

[0036] The second frustum 112, serving as the middle section, connects the first cone 111 and the cylinder 113. Its generatrix angle β is smaller than the generatrix angle θ of the first cone 111, and the angle β ranges from 10° to 15°, with values ​​including but not limited to 10°, 11°, 12°, 13°, 14°, and 15°. It is primarily used to carry and receive the sample mixed with the first reagent, as well as the second reagent in channel 21. The second frustum 112, with its gentle cone angle, acts as an expanded mixing zone, mainly used to carry the sample and ensure thorough mixing and reaction with the reagents from the upper and lower sections.

[0037] The upper cylindrical body 113 has an internal groove / protrusion or thread that mates with the sealing cap 12. This is to allow the detection unit 10 and the pipetting unit 20 to form a sealed chamber. The cylinder 113 serves as a standard interface, connecting with the pipetting unit 20 through its engagement with the sealing cap 12, together forming a sealed chamber for detection.

[0038] The detection cup 11 has a volume of 180-200µL, and its inner surface is treated with a low-adsorption process. Volumes include, but are not limited to, 180µL, 182µL, 184µL, 186µL, 188µL, 190µL, 192µL, 194µL, 196µL, 198µL, and 200µL. The detection cup 11 is made of high-transmittance medical-grade polypropylene (PP) or cyclic olefin copolymer (COC / COP), with a transmittance ≥90%.

[0039] The total volume of the reagents in the test cup 11 is larger than the volume of the first reagent it pre-loads, in order to receive samples such as nucleic acids.

[0040] A sealing cap 12 is also provided on the test cup 11 for sealing cooperation, and the sealing cap 12 is detachably connected to the test cup 11. The sealing cap 12 can be connected to the test cup 11 through a bayonet fit or a threaded fit. The pipetting unit 20 and the sealing cap 12 are integrally injection molded, such as... Figure 6 As shown; or, after separate assembly, it is fixed to the lower surface of the sealing cover 12, as shown. Figure 4 As shown. Regardless of how the sealing cap and the pipetting unit 20 are connected, the final example structure is as follows. Figure 2-3 As shown, the purpose is to connect the pipetting unit 20 and the detection unit 10 to the detection cup 11 through the channel 21 and the sealing cap 12, forming a sealed communication chamber, thereby preventing aerosol leakage during the amplification process.

[0041] When the sealing cap 12 and the pipetting unit 20 are assembled separately, the center of the sealing cap 12 has an elastic silicone pad or membrane valve that matches the outer diameter of the channel 21. Once punctured, it fits snugly against the channel 21, forming a closed, connected chamber. Sample nucleic acid is injected and reconstituted with lyophilized reagents. The sample nucleic acid is injected into the test cup 11 by puncturing the elastic silicone pad or membrane valve of the sealing cap 12 with a volume of 10-50 µL. The elastic silicone pad / membrane valve of the sealing cap 12 automatically closes after puncture, maintaining airtightness. The bottom of the test cup 11 is pre-loaded with lyophilized nucleic acid amplification microspheres, which instantly reconstitute upon contact with liquid. This design avoids opening the cap, blocking aerosol contamination sources; moreover, the lyophilized reagents are stable at room temperature, eliminating the need for cold chain transportation and on-site preparation.

[0042] like Figure 7-8 As shown, the volume of the channel in the pipetting unit 20 is 10-100µL, including but not limited to 10µL, 15µL, 20µL, 25µL, 30µL, 35µL, 40µL, 45µL, 50µL, 55µL, 60µL, 65µL, 70µL, 75µL, 80µL, 85µL, 90µL, 95µL, and 100µL.

[0043] The pipetting unit 20 also includes a second vesicle 23. The first vesicle 22 and the second vesicle 23 are interconnected and communicate with each other through a short microchannel. They have the same structure, both being rectangular, sealed capsules. The first vesicle 22 and the second vesicle 23 are both located on the side of the channel 21 and on the same side. The first vesicle 22 and the second vesicle 23 are integrally formed with the channel 21, which facilitates processing and operation. The first vesicle 22 is the active operating area, which provides power when squeezed to push the liquid in the channel into the detection cup 11. The second vesicle 23 is a buffer / collection area located at the end of the channel; it can accommodate excess liquid that may flow back from the channel, ensuring that only a predetermined volume of liquid enters the detection cup, thereby ensuring the accuracy of the transfer volume.

[0044] Furthermore, one end of the second vesicle 23 is connected to the channel 21, and the other end is connected to the first vesicle 22; the second vesicle 23 is located at the end of the channel 21 away from the detection cup 11, and the first vesicle 22 is located above the second vesicle 23. The pipetting unit 20 adopts a double vesicle structure, which can ensure that the liquid can be accurately and controllably transferred quantitatively and prevent backflow or overflow; this vertically arranged structure of the first vesicle 22 and the second vesicle 23 helps to utilize gravity or pressure difference to make the liquid flow orderly between the vesicle and the channel.

[0045] Figure 7 This is an example diagram of the second reagent in pipetting unit 20 before transfer, and an example diagram of it after transfer is shown below. Figure 8 As shown.

[0046] The channel 21, pre-filled with a second reagent such as a fluorescent dye or enzyme mixture, is assembled with the top of the sealing cap 12. The channel 21 contains a pre-quantitative amount of the second reagent (10-100 µL), with any excess reagent temporarily stored in the second vesicle 23. The channel 21 is locked to the sealing cap 12 via a snap-fit ​​or integral molding. The fixed volume of the channel 21 enables precise quantification; moreover, the assembly forms a completely sealed system, preventing external contamination.

[0047] The entire consumable is placed in a thermostatic device, such as a LAMP / RPA instrument, and amplified at 61-65℃ for 15-30 minutes for isothermal amplification reaction. The high-transmittance material (transmittance ≥90%) and conical bottom design of the test cup 11 ensure uniform temperature conduction; moreover, the airtight chamber of the sealed cap 12 prevents reaction vapor leakage. The conical bottom reduces thermal mass and accelerates temperature equilibrium; and the fully enclosed system avoids false positives caused by leakage of amplification products.

[0048] In the constant temperature device, the drive module can squeeze the first vesicle 22 through its squeezing head, causing the quantitative second reagent in the channel 21 to flow into the detection cup 11, thus transferring the second reagent. The squeezing head is a common structure in the art, generally an arc-shaped silicone contact, with a pressure of 0.1-0.5N to avoid vesicle rupture (see attached diagram). If liquid overflows and rises in the channel 21 during the squeezing of the first vesicle 22, the second vesicle 23 can be used to collect the overflowing liquid, ensuring that the transferred volume does not overflow. The entire squeezing process is completed within a closed system, achieving "zero-opening" reagent addition; moreover, no manual intervention is required, and the device automatically triggers the transfer action.

[0049] In the actual testing process, a dedicated optical detection module (a common detection method in this field) is also provided, which directly collects fluorescence / colorimetric signals from directly below the conical bottom of the detection cup 11 through optical signals and reads the corresponding results.

[0050] The detection cup 11, made of a high-transmittance material, and its conical bottom surface with optical focusing design, can increase the detection signal intensity by ≥20%. Furthermore, there is no need to open the lid or transfer the mixed solution; the liquid inside the detection cup 11 is directly irradiated by the optical detection module, and the detection result is read directly. It is convenient, safe, and offers high reading accuracy, without any other human interference.

[0051] An isothermal nucleic acid amplification and detection device, adapted to the quantitative transfer consumables described above.

[0052] This application discloses a isothermal nucleic acid amplification and detection quantitative transfer consumable and detection device, which can accurately transfer liquid quantitatively. The entire detection process requires no opening of the lid, preventing aerosol contamination and significantly reducing false positives. The channel has a fixed volume, resulting in small liquid transfer errors and accurate quantification. The detection cup can be pre-filled with lyophilized reagents, stored and transported at room temperature, and used immediately upon opening, eliminating the need for a cold chain and precise liquid transfer. The conical bottom and high light transmittance design enhance the fluorescence signal by >20%, making it suitable for low-copy detection. It is compatible with existing standard tubular reaction systems, and single-use design avoids cross-contamination.

[0053] The embodiments of this application have been described in detail above. These descriptions are merely preferred embodiments and should not be construed as limiting the scope of this application. All equivalent variations and modifications made within the scope of this application should still fall within the patent coverage of this application.

Claims

1. A quantitative transfer consumable for isothermal nucleic acid amplification detection, characterized in that, include: The detection unit is equipped with at least one detection cup that can be pre-loaded with the first reagent and can receive the sample; The pipetting unit has a channel for pre-storing a second reagent and at least one first vesicle communicating with the channel; The pipetting unit and the detection unit are connected to the detection cup through the channel to form a closed and connected chamber; and when the first vesicle is squeezed, the second reagent can enter the detection cup through the channel.

2. The isothermal nucleic acid amplification and quantitative transfer consumable according to claim 1, characterized in that, The detection cup has a conical structure, comprising a lower first cone, a middle second frustum, and an upper cylinder; the generatrix angle of the first cone is greater than the generatrix angle of the second frustum.

3. The isothermal nucleic acid amplification and quantitative transfer consumable according to claim 2, characterized in that, The volume of the detection cup is greater than the volume of the first reagent; and when the first vesicle is squeezed, the channel is inserted into the first cone, so that the second reagent is squeezed into the first cone.

4. A quantitative transfer consumable for isothermal nucleic acid amplification detection according to any one of claims 1-3, characterized in that, The test cup is also provided with a sealing cap that seals with it, and the sealing cap is detachably connected to the test cup.

5. The isothermal nucleic acid amplification and quantitative transfer consumable according to claim 4, characterized in that, The channel is integrally injection molded with the sealing cover, or it is assembled separately and then fixed to the lower surface of the sealing cover.

6. A quantitative transfer consumable for isothermal nucleic acid amplification detection according to any one of claims 1-3 and 5, characterized in that, The pipetting unit also includes a second vesicle, and the first vesicle and the second vesicle are interconnected.

7. The isothermal nucleic acid amplification and quantitative transfer consumable according to claim 6, characterized in that, The first vesicle and the second vesicle are both located on the same side of the channel; and the first vesicle, the second vesicle, and the channel are integrally connected.

8. The isothermal nucleic acid amplification and quantitative transfer consumable according to claim 7, characterized in that, One end of the second vesicle is connected to the channel, and the other end is connected to the first vesicle.

9. A quantitative transfer consumable for isothermal nucleic acid amplification detection according to claim 7 or 8, characterized in that, The second vesicle is located at the end of the channel away from the detection cup, and the first vesicle is located above the second vesicle.

10. An isothermal nucleic acid amplification and detection device, characterized in that, Adapted to the quantitative transfer consumables as described in any one of claims 1-9.

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