A detection kit for inactivating a COVID-19 antigen reagent

Abstract

The present invention relates to a detection kit for inactivating a new coronavirus antigen reagent, belonging to the technical field of biotechnology. It includes a housing, in which a colloidal gold chromatography reagent strip and an inactivating reaction solution are provided. The composition components of the inactivating reaction solution include guanidine isothiocyanate with a concentration of 0.28 mol / L, Tris-HCl with a concentration of 10 mmol / L, and disodium ethylenediaminetetraacetate with a concentration of 1 mmol / L. The detection samples of the present invention are diverse. On the premise of not affecting the detection sensitivity and specificity, an inactivating reaction solution capable of safely and efficiently inactivating virus samples is provided, which can quickly and effectively inactivate the new coronavirus after detection using an antigen reagent, realizing the integration of detection and inactivation; the inactivating reaction solution is safe and non-toxic, easy to use, and can minimize the risk of medical staff and testers being infected with the new coronavirus.

Description

Technical Field

[0001] The present invention relates to the technical field of biotechnology. Specifically, it relates to a detection kit for inactivating a new coronavirus antigen reagent. Background Art

[0002] The new coronavirus is a type of non-cellular microorganism composed of a protein coat and nucleic acid (RNA), and can mutate continuously by adapting to the host during the replication process. It usually causes symptoms such as headache, cough, nasal congestion, fever, and difficulty breathing in humans. Some cases will develop into pneumonia and even cause death. Due to the strong infectivity of this virus, improper handling after antigen detection will cause great harm to human health and the public environment.

[0003] Currently, the detection of the new coronavirus mainly relies on nucleic acid detection and antigen detection. Nucleic acid detection technology requires the use of complex precision instruments, with cumbersome operations and long time consumption. While antigen detection technology can complete the detection of the new coronavirus through a colloidal gold chromatography reagent strip, with simple operation and short time consumption, and can achieve home self-testing. The new coronavirus usually requires the collection of nasal swabs, throat swabs, sputum, or alveolar lavage fluid from patients for detection. However, due to the very strong infectivity of the new coronavirus, it poses a great risk of infection to medical staff and testers. Therefore, after the collection of new coronavirus samples, immediate inactivation treatment is required, which can effectively reduce the risk of infection for medical staff and testers. However, when the sample is inactivated, although the virus loses its original biological activity, its detection markers (RNA and proteins with spatial structures) will also be damaged and difficult to detect. Therefore, on the premise of effectively reducing the risk of infection for medical staff and testers, achieving accurate detection of the new coronavirus has become an urgent key issue. Summary of the Invention

[0004] The purpose of the present invention is to provide a detection kit for inactivating a new coronavirus antigen reagent, which has a variety of detection sample types. On the premise of not affecting the detection sensitivity and specificity, it provides an inactivating reaction solution that can safely and efficiently inactivate virus samples, and can quickly and effectively inactivate the new coronavirus after detection using an antigen reagent, realizing the integration of detection and inactivation; the inactivating reaction solution is safe, non-toxic, easy to use, and can minimize the risk of medical staff and testers being infected with the new coronavirus.

[0005] To achieve the above purpose, the technical solution adopted by the present invention is as follows: A detection kit for inactivating a new coronavirus antigen reagent, including a housing, in which a colloidal gold chromatography reagent strip and an inactivating reaction solution are provided. The composition components of the inactivating reaction solution include guanidine isothiocyanate with a concentration of 0.28 mol / L, Tris-HCl with a concentration of 10 mmol / L, and disodium ethylenediaminetetraacetate with a concentration of 1 mmol / L.

[0006] As a further improvement of the above technical solution:

[0007] The preparation method of the inactivated reaction solution includes the following steps: Weigh 33 g of guanidine isothiocyanate, 1.21 g of Tris-HCl, and 0.34 g of disodium ethylenediaminetetraacetate in a 1 L beaker for volume fixation, add 800 mL of sterile water for dissolution, mix well, pour it into a 1 L volumetric flask after dissolution, take about 200 mL of sterile water and wash the beaker three times and pour it into the volumetric flask, adjust the pH value of the solution to be between 7.0 - 7.4, add sterile water to fix the volume to 1 L, and perform high-temperature sterilization with an autoclave to obtain the inactivated reaction solution.

[0008] The pH value of the sterile water is 7.2 ± 0.2.

[0009] A partition is provided inside the housing, and the inside of the housing is divided into a reaction chamber and an inactivated reaction solution storage chamber by the partition. The colloidal gold chromatography reagent strip is arranged in the reaction chamber, and the inactivated reaction solution is injected into the inactivated reaction solution storage chamber.

[0010] An inactivated reaction switch is provided on the partition.

[0011] The colloidal gold chromatography reagent strip includes a sample pad, a conjugate pad, a nitrocellulose membrane, a water absorption pad, and a bottom plate. The sample pad, the conjugate pad, the nitrocellulose membrane, and the water absorption pad are sequentially connected in the horizontal direction and arranged on the bottom plate.

[0012] The conjugate pad is conjugated with streptavidin-biotin amplified colloidal gold nanoparticles.

[0013] The nitrocellulose membrane is provided with a detection line coated with a new coronavirus antigen capture antibody and a quality control line coated with an internal reference capture antibody, and the detection line and the quality control line are arranged in sequence along the chromatography direction.

[0014] The housing is provided with a sample addition hole and a detection window, and the reaction chamber is communicated with the sample addition hole and the detection window.

[0015] The sample pad is located at the sample addition hole, and the sample addition hole gradually narrows from the outer end to the inner end, and the detection line and the quality control line are located at the detection window.

[0016] The present invention adopts the above technical solutions and has the following advantages:

[0017] 1. On the basis of not affecting the detection sensitivity and specificity of the antigen, it can quickly inactivate the detected new coronavirus, realizing the integration of detection and inactivation, and can effectively avoid the risk of secondary transmission of the new coronavirus.

[0018] 2. Low biosafety risk: Inactivate the virus by adding guanidine salts, which takes effect quickly, is non-toxic and harmless, and will not pose a biosafety risk, thus protecting medical staff and testers from the risk of COVID-19 infection to the greatest extent.

[0019] 3. Fast and simple: The test kit that can inactivate COVID-19 antigen reagents is easy to operate, does not rely on sophisticated nucleic acid extractors and fluorescence quantitative PCR instruments, has a fast sample detection speed, and the test results can be observed with the naked eye, enabling home self-testing and inactivation treatment of COVID-19 virus.

[0020] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. Description of the Drawings

[0021] Figure 1 It is a schematic structural diagram of a test kit that can inactivate COVID-19 antigen reagents in an embodiment of the present invention;

[0022] Figure 2 It is a schematic structural diagram inside the housing 1 of a test kit that can inactivate COVID-19 antigen reagents in an embodiment of the present invention;

[0023] Figure 3 It is a schematic structural diagram of a colloidal gold chromatography reagent strip in an embodiment of the present invention.

[0024] In the figure, 1 - housing; 2 - sample adding hole; 3 - detection window; 4 - test line; 5 - quality control line; 6 - bottom plate; 7 - nitrocellulose membrane; 8 - inactivation reaction switch; 9 - partition; 10 - sample pad; 11 - coupling pad; 12 - absorbent pad; 13 - colloidal gold chromatography reagent strip; 14 - reaction chamber; 15 - inactivation reaction liquid storage chamber. Specific Embodiments

[0025] The implementation schemes of the present invention will be described in detail below in conjunction with the embodiments. For the conditions not specified in the embodiments, they are carried out according to the conventional conditions or the conditions recommended by the manufacturer. For the reagents or instruments not specified by the manufacturer, they are all conventional products that can be obtained through market purchase.

[0026] The described embodiments are only a part of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts fall within the scope of protection of the present invention.

[0027] Example 1:

[0028] As Figures 1-3As shown in the figure, a detection kit for inactivating a new coronavirus antigen reagent includes a housing 1. Inside the housing 1, there is a colloidal gold chromatography reagent strip 13 and an inactivating reaction solution. A partition 9 is provided inside the housing 1, and the partition 9 divides the interior of the housing 1 into a reaction chamber 14 and an inactivating reaction solution storage chamber 15. The colloidal gold chromatography reagent strip 13 is arranged in the reaction chamber 14, and the inactivating reaction solution is injected into the inactivating reaction solution storage chamber 15. An inactivating reaction switch 8 is provided on the partition 9.

[0029] The colloidal gold chromatography reagent strip 13 includes a sample pad 10, a conjugate pad 11, a nitrocellulose membrane 7, an absorbent pad 12, and a base plate 6. The sample pad 10, the conjugate pad 11, the nitrocellulose membrane 7, and the absorbent pad 12 are sequentially connected in the horizontal direction and arranged on the base plate 6. Streptavidin-biotin amplified colloidal gold nanoparticles are conjugated on the conjugate pad 11. A detection line 4 coated with a new coronavirus antigen capture antibody and a quality control line 5 coated with an internal reference capture antibody are provided on the nitrocellulose membrane 7. The detection line 4 and the quality control line 5 are arranged in sequence along the chromatography direction.

[0030] A sample adding hole 2 and a detection window 3 are provided on the housing 1. The reaction chamber 14 communicates with the sample adding hole 2 and the detection window 3. The sample pad 10 is located at the sample adding hole 2. The sample adding hole 2 gradually narrows from the outer end to the inner end. The detection line 4 and the quality control line 5 are located at the detection window 3.

[0031] The composition components of the inactivating reaction solution include guanidine isothiocyanate with a concentration of 0.28 mol / L, Tris-HCl with a concentration of 10 mmol / L, and disodium ethylenediaminetetraacetate with a concentration of 1 mmol / L. The main function of guanidine isothiocyanate is to act as a protein denaturant and is also used for denaturing and lysing viral RNA. The main function of Tris-HCl is to provide a suitable virus lysis environment. The main function of disodium ethylenediaminetetraacetate is to chelate divalent metal ions. The solvent of the inactivating reaction solution is sterile water, and the pH value is 7.2 ± 0.2.

[0032] The preparation method of the inactivating reaction solution includes the following steps: Weigh 33 g of guanidine isothiocyanate, 1.21 g of Tris-HCl, and 0.34 g of disodium ethylenediaminetetraacetate into a 1 L beaker for volume fixing, add 800 mL of sterile water for dissolution, stir well with a glass rod until fully mixed. After dissolution, pour it into a 1 L volumetric flask. Take about 200 mL of sterile water and wash the beaker three times and pour it into the volumetric flask. Adjust the pH value of the solution to be between 7.0 - 7.4, add sterile water to make the volume up to 1 L, and perform high-temperature sterilization with an autoclave to obtain the inactivating reaction solution.

[0033] The applicable sample of the detection kit for inactivating a new coronavirus antigen reagent in the present invention is a respiratory sample, including nasal swabs, throat swabs, sputum, or bronchoalveolar lavage fluid, and the object to be inactivated is a new coronavirus sample.

[0034] The detection kit for inactivating the new coronavirus antigen reagent of the present invention can not only achieve the specific and sensitive detection of the new coronavirus, but also completely inactivate the new coronavirus after the detection is completed, effectively reducing the risk of infection for medical staff and testers, and at the same time can avoid causing harm to the environment, and avoid the secondary transmission and infection of the new coronavirus from the source.

[0035] Example 2:

[0036] An experimental group and a control group were set up. In the experimental group, 500 μL of lentivirus with a titer of 2×10 8 TU / mL of high-brightness green fluorescent protein was mixed with 500 μL of the inactivating reaction solution prepared in Example 1. After mixing for 5 minutes, the mixture was taken to infect Vero cells. The infected Vero cells were cultured for 3 days, and the expression of green fluorescent protein was observed under a fluorescence microscope to verify the virus activity. In the control group, lentivirus with a titer of 10 8 TU / mL of high-brightness green fluorescent protein was directly used to infect Vero cells. The infected Vero cells were cultured for 3 days, and the expression of green fluorescent protein was observed under a fluorescence microscope to verify the virus infection activity.

[0037] The verification results were as follows: The expression result of green fluorescent protein in the experimental group under the fluorescence microscope did not show green, indicating that the virus had been completely inactivated by the inactivating reaction solution and lost its ability to infect cells; the control group produced obvious green fluorescence under the fluorescence microscope, indicating that the virus had strong infectivity.

[0038] The experimental results showed that the lentivirus was completely inactivated after being treated in the inactivating reaction solution of the present invention for 5 minutes, indicating that the inactivating reaction solution of the present invention had a good inactivating effect on the virus.

[0039] Comparative Example 1:

[0040] Weigh 1.21 g of Tris-HCl and 0.34 g of disodium ethylenediaminetetraacetate respectively, add 800 mL of sterile water for dissolution, stir well with a glass rod until fully mixed, pour into a 1 L volumetric flask for volume fixation after dissolution, take about 200 mL of sterile water and wash the beaker three times and pour it into the volumetric flask, adjust the pH of the solution to between 7.0 and 7.4, add sterile water to make up the volume to 1 L, and sterilize it at high temperature with an autoclave to obtain the inactivating reaction solution. The concentration of Tris-HCl in the inactivating reaction solution is 10 mmol / L, and the concentration of disodium ethylenediaminetetraacetate is 1 mmol / L.

[0041] In this comparative example, the guanidine isothiocyanate component was removed, and it was found that the lentivirus could not be completely inactivated and still had strong virus activity after 3 days of cell culture, indicating that removing guanidine isothiocyanate from the inactivated reaction solution would lead to the risk of COVID-19 virus infection.

[0042] Comparative Example 2:

[0043] In this comparative example, the concentration of guanidine isothiocyanate in the inactivated reaction solution was 1 mol / L, and the other components and their concentrations were the same as those in Example 1. It was placed in a refrigerator at 4°C for 24 h to simulate the low-temperature transportation environment, and a small amount of crystallization was found, indicating that the inactivated reaction solution containing a high concentration of guanidine isothiocyanate had poor stability; while the inactivated reaction solution in Example 1 was placed in a refrigerator at 4°C for 24 h or longer (3 months), and no crystallization was found, indicating that the inactivated reaction solution in Example 1 had good stability.

[0044] In summary, a detection kit for an inactivated COVID-19 antigen reagent of the present invention is safe, non-toxic, convenient and fast to use, and can be widely used for inactivating virus samples under normal temperature conditions in hospitals, families, research institutes, etc. After the collected COVID-19 virus sample is detected by an antigen detection reagent strip, the inactivated reaction switch 8 is turned on, and the inactivated reaction solution automatically enters the reaction chamber 14 containing the COVID-19 virus sample. By slowly shaking the housing 1, the COVID-19 virus can be completely inactivated, which can not only minimize the risk of secondary infection of medical staff and testers, but also reduce the harm to the environment.

[0045] Finally, it should be noted that the above are only preferred embodiments of the present invention and are not used to limit the present invention. Examples of implementation methods are given, and the parts not described in detail are all common general knowledge of those of ordinary skill in the art. For those skilled in the art, they can still modify the technical solutions recorded in the foregoing embodiments or perform equivalent replacements on some of the technical features. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A detection kit for inactivating a new coronavirus antigen reagent, characterized in that: It includes a housing, inside which there is a colloidal gold chromatography reagent strip and an inactivated reaction solution. The components of the inactivated reaction solution include guanidine isothiocyanate with a concentration of 0.28 mol / L, Tris-HCl with a concentration of 10 mmol / L, and disodium ethylenediaminetetraacetate with a concentration of 1 mmol / L. The preparation method of the inactivated reaction solution includes the following steps: Weigh 33 g of guanidine isothiocyanate, 1.21 g of Tris-HCl, and 0.34 g of disodium ethylenediaminetetraacetate into a 1 L beaker for volume fixation, add 800 mL of sterile water for dissolution, mix well, pour it into a 1 L volumetric flask after dissolution, take 200 mL of sterile water to wash the beaker three times and pour it into the volumetric flask, adjust the pH value of the solution to be between 7.0 - 7.4, add sterile water to fix the volume to 1 L, and perform high-temperature sterilization with an autoclave to obtain the inactivated reaction solution.

2. The detection kit for inactivating the novel coronavirus antigen reagent according to claim 1, wherein: The pH value of the sterile water is 7.2 ± 0.

2.

3. A detection kit for inactivating a new coronavirus antigen reagent according to claim 1, characterized in that: There is a partition inside the housing, which divides the inside of the housing into a reaction chamber and an inactivated reaction solution storage chamber. The colloidal gold chromatography reagent strip is arranged in the reaction chamber, and the inactivated reaction solution is injected into the inactivated reaction solution storage chamber.

4. A detection kit for an inactivatable new coronavirus antigen reagent according to claim 3, characterized in that: There is an inactivated reaction switch on the partition.

5. A detection kit for inactivating a new coronavirus antigen reagent according to claim 1, characterized in that: The colloidal gold chromatography reagent strip includes a sample pad, a conjugate pad, a nitrocellulose membrane, an absorbent pad, and a bottom plate. The sample pad, the conjugate pad, the nitrocellulose membrane, and the absorbent pad are sequentially connected in the horizontal direction and arranged on the bottom plate.

6. The detection kit for inactivating a new coronavirus antigen reagent according to claim 5, characterized in that: The conjugate pad is conjugated with streptavidin-biotin amplified colloidal gold nanoparticles.

7. The detection kit for inactivating a new coronavirus antigen reagent according to claim 6, wherein: There are a detection line coated with a new coronavirus antigen capture antibody and a quality control line coated with an internal reference capture antibody on the nitrocellulose membrane, and the detection line and the quality control line are arranged in sequence along the chromatography direction.

8. A detection kit for inactivating a novel coronavirus antigen reagent according to claim 7, characterized in that: There are a sample addition hole and a detection window on the housing, and the reaction chamber communicates with the sample addition hole and the detection window.

9. The detection kit for inactivating a novel coronavirus antigen reagent according to claim 8, characterized in that: The sample pad is located at the sample addition hole, and the sample addition hole gradually narrows from the outer end to the inner end. The detection line and the quality control line are located at the detection window.

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