Detection kit of novel coronavirus antigen and application, detection method of novel coronavirus antigen

Abstract

The application provides a detection kit for a novel coronavirus antigen, comprising a capture antibody, a biotin-labeled detection antibody, a SARS-CoV2 N protein standard sample, an enzyme-labeled avidin, a chromogenic solution and a termination solution, wherein the capture antibody comprises an anti-SARS-CoV2 N protein capture antibody, the sequence of the heavy chain variable region of the anti-SARS-CoV2 N protein capture antibody comprises an amino acid sequence as shown in SEQ ID NO:1, and the sequence of the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:2. The detection kit can qualitatively and quantitatively detect the novel coronavirus, has a low detection limit and a wide linear range, can accurately and efficiently detect the novel coronavirus in an early stage, and is beneficial to the prevention and control of the novel coronavirus pneumonia.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medical biology, in particular to a novel coronavirus antigen detection kit and application, and a novel coronavirus antigen detection method. BACKGROUND

[0002] The novel coronavirus (SARS-CoV-2) causes COVID-19, which is the seventh known coronavirus that can infect humans. The detection of the novel coronavirus has a crucial impact on the prevention and control of COVID-19.

[0003] Currently, the detection methods of the novel coronavirus mainly include serum antibody detection and viral nucleic acid detection. Virus-specific antibodies generally appear about 5-15 days after the virus invades the human body, so early serum antibody detection is not possible, and there are serious false negative and false positive results in serum antibody detection. At the same time, the quality of the novel coronavirus nucleic acid detection kit is uneven, the reliability of nucleic acid detection is poor, the nucleic acid detection rate is low, and many cases require repeated detection for 2-3 times, but there are still many false positive and false negative results. Therefore, how to improve the early detection rate and accuracy of the novel coronavirus is of great significance for the prevention and control of COVID-19. SUMMARY

[0004] Therefore, the present application provides a novel coronavirus antigen detection kit, which has a low detection limit, can detect the novel coronavirus in the early stage of infection, has high detection accuracy, can realize rapid detection, and is conducive to the prevention and control of COVID-19.

[0005] In a first aspect, the present application provides a novel coronavirus antigen detection kit, which comprises a capture antibody, a biotin-labeled detection antibody, a SARS-CoV2 N protein standard, an enzyme-labeled avidin, a chromogenic solution and a termination solution, wherein the capture antibody comprises an anti-SARS-CoV2 N protein capture antibody, the sequence of the heavy chain variable region of the anti-SARS-CoV2 N protein capture antibody comprises an amino acid sequence as shown in SEQ ID NO: 1, and the sequence of the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 2.

[0006] The detection kit for the novel coronavirus antigen provided in the application can qualitatively and quantitatively detect the novel coronavirus; based on the antigen-antibody interaction, the capture antibody binds with the sample to be detected, and the sample to be detected binds with the detection antibody, and the enzyme-labeled avidin and biotin can specifically bind, thereby forming a "capture antibody-sample to be detected-biotin-labeled detection antibody-enzyme-labeled avidin" complex, and through the color development reaction, the sample to be detected can be qualitatively detected; further, by using the anti-SARS-CoV2 N protein capture antibody with a specific sequence as the capture antibody, the capture antibody can specifically and efficiently bind with the antigen, reduce the detection limit of the novel coronavirus, improve the linear detection range, and be more conducive to the use of the detection kit.

[0007] Optionally, the detection antibody is a rabbit anti-SARS-CoV2 N protein antibody with a product number of 40143-R040 from Beijing Yiqiao God Science and Technology Co., Ltd.

[0008] Optionally, the SARS-CoV2 N protein standard product is an N protein standard product with a product number of B232004 from Taizhou Baiying Biological Technology Co., Ltd.

[0009] Optionally, the detection kit for the novel coronavirus antigen has a minimum detection limit of 46.8 pg / mL. The detection kit for the novel coronavirus antigen provided in the application has a low detection limit, and the novel coronavirus can be detected when the novel coronavirus reaches 46.8 pg / mL, thereby being conducive to the prevention and control of the novel coronavirus pneumonia.

[0010] Optionally, the detection kit for the novel coronavirus antigen has a linear range of 93.74 pg / mL-1200 pg / mL. The detection kit for the novel coronavirus antigen provided in the application has a wide linear detection range and high accuracy, and can be applied to the detection of the novel coronavirus in different scenarios.

[0011] Optionally, the detection kit further comprises a washing liquid, and the washing liquid is a phosphate buffer solution containing Tween.

[0012] Optionally, the detection kit further comprises a blocking liquid, and the blocking liquid is a phosphate buffer solution containing bovine serum albumin and / or skimmed milk. The use of the above blocking liquid can effectively block the binding sites in the detection hole that are not coated with the capture antibody, thereby improving the accuracy of the detection.

[0013] Optionally, the enzyme-labeled avidin comprises peroxidase-labeled avidin. Further, the enzyme-labeled avidin comprises horseradish peroxidase-labeled avidin. Further, the enzyme-labeled avidin comprises horseradish peroxidase-labeled streptavidin.

[0014] Optionally, the color developing solution comprises a 3,3',5,5'-tetramethylbenzidine (TMB) solution. Under the action of peroxidase, the color of the TMB solution changes, so that the novel coronavirus in the sample to be detected can be qualitatively and quantitatively detected according to whether the color of the TMB solution changes and the absorbance of the solution before and after the color change.

[0015] Optionally, the termination solution comprises a sulfuric acid solution. By adding the sulfuric acid solution, the activity of the enzyme molecules in the enzyme-labeled avidin can be destroyed, so that the color developing reaction between the enzyme molecules and the color developing solution is stopped, and the accuracy of the detection result is ensured.

[0016] The detection limit of the novel coronavirus antigen detection kit provided in the application is low, which can detect the novel coronavirus in the early stage of infection, so as to effectively prevent and control the novel coronavirus pneumonia. At the same time, the detection kit can accurately detect the novel coronavirus, avoid false positive and false negative phenomena, and the detection is rapid, which is beneficial to its application.

[0017] In a second aspect, the application provides the use of the novel coronavirus antigen detection kit of the first aspect in the detection of a novel coronavirus antigen for a purpose other than disease diagnosis and / or treatment.

[0018] The novel coronavirus antigen detection kit provided in the application can be used in the detection of a novel coronavirus antigen for a purpose other than disease diagnosis and / or treatment, such as detecting a novel coronavirus antigen in the environment, so as to determine whether the environment is safe, which is beneficial to the prevention and control of the novel coronavirus pneumonia.

[0019] In a third aspect, the application provides a method for detecting a novel coronavirus antigen for a purpose other than disease diagnosis and / or treatment, comprising detecting a novel coronavirus antigen by using the novel coronavirus antigen detection kit of the first aspect.

[0020] Optionally, the detection method comprises:

[0021] The capture antibody is dispersed in a phosphate buffer to obtain a capture antibody solution, and the biotin-labeled detection antibody is dispersed in a phosphate buffer to obtain a detection antibody solution;

[0022] The capture antibody solution is added to the detection hole so that the capture antibody is coated in the detection hole;

[0023] The pretreated sample to be detected is added to the detection hole for incubation, and then washed;

[0024] The detection antibody solution is added to the detection hole for incubation, and then washed;

[0025] After adding the enzyme-labeled avidin to the detection hole, a color developing solution is added for color developing reaction, and then a termination solution is added to terminate the color developing reaction;

[0026] Qualitative detection is performed according to the color change of the detection hole, and / or the OD 450 value of the detection hole is measured and compared with a standard curve to obtain the concentration of the novel coronavirus antigen in the sample to be detected.

[0027] Further, the preparation of the standard curve includes:

[0028] The capture antibody is coated in a plurality of detection holes;

[0029] The SARS-CoV2 N protein standard is diluted to form a plurality of SARS-CoV2 N protein solutions with different concentrations, and the plurality of SARS-CoV2 N protein solutions with different concentrations are respectively added to different detection holes, and then washing is performed;

[0030] The detection antibody solution is added to each detection hole for incubation, and then washing is performed;

[0031] After adding the enzyme-labeled avidin to the detection hole, a color developing solution is added for color developing reaction, and then a termination solution is added to terminate the color developing reaction;

[0032] The OD 450 values of a plurality of detection holes are measured, and the standard curve is obtained according to the concentration of the SARS-CoV2 N protein solution and the OD 450 value.

[0033] Further, the concentration ratio of the SARS-CoV2 N protein solution to the detection antibody is 4.68x10 -5 -1.2x10 -2 -1.2x10 -2 By using the above concentration ratio, the SARS-CoV2 N protein and the detection antibody can be efficiently, rapidly and specifically combined, the minimum detection limit is reduced, and the novel coronavirus can be effectively detected when the concentration of the novel coronavirus is low in the early stage.

[0034] Further, the concentration of the color developing solution is 0.01%-0.2%, and the concentration of the termination solution is 1M-4M.

[0035] The novel coronavirus antigen detection method provided in the application is simple in operation, short in detection time, convenient in sampling, reduces the probability of false positive and false negative, and improves the detection accuracy. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1A flow chart of the detection method of the novel coronavirus antigen provided for an embodiment of the present application.

[0037] Figure 2 A flow chart of the preparation of the standard curve in the detection method of the novel coronavirus antigen provided for an embodiment of the present application.

[0038] Figure 3 A standard curve chart in the detection method of the novel coronavirus antigen provided for an embodiment of the present application. DETAILED DESCRIPTION

[0039] The following describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, which are also considered within the scope of protection of the present application.

[0040] The present application provides a novel coronavirus antigen detection kit, comprising a capture antibody, a biotin-labeled detection antibody, a SARS-CoV2 N protein standard, an enzyme-labeled avidin, a chromogenic solution and a termination solution, the capture antibody comprising an anti-SARS-CoV2 N protein capture antibody, the sequence of the heavy chain variable region of the anti-SARS-CoV2 N protein capture antibody comprising the amino acid sequence shown in SEQ ID NO: 1, and the sequence of the light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 2.

[0041] In related technologies, when serum antibodies are used for novel coronavirus detection, the production of antibodies in serum requires time, so that effective detection cannot be performed in the early stage, and false positive and false negative phenomena are serious; when the novel coronavirus is detected by viral nucleic acid, a complex process such as nucleic acid extraction is required, so that the extraction process is easily contaminated, reducing the detection speed and accuracy. The detection kit provided by the present application is for the nucleocapsid protein (N protein) of the novel coronavirus, which can be produced in the early stage of infection of the patient with the novel coronavirus, thereby facilitating early detection of the novel coronavirus, and is of great significance for timely diagnosis, treatment of the novel coronavirus patient and prevention and control of the novel coronavirus. Further, the anti-SARS-CoV2 N protein capture antibody with a specific sequence used in the detection kit provided by the present application can efficiently and specifically bind to the antigen, which is conducive to the quantitative detection of the novel coronavirus at low concentration, and plays an auxiliary role in the assessment of the condition of the novel coronavirus patient, thereby improving the cure rate of the novel coronavirus patient.

[0042] In the embodiments of the present application, the detection antibody is rabbit anti-SARS-CoV2 N protein antibody with product number 40143-R040 from Beijing Yiqiao God Science and Technology Co., Ltd. In the embodiments of the present application, the SARS-CoV2 N protein standard is N protein standard with product number B232004 from Taizhou Baiying Biological Technology Co., Ltd. By using the above detection antibody and SARS-CoV2 N protein standard, in combination with the anti-SARS-CoV2 N protein capture antibody with a specific sequence, the detection limit of the novel coronavirus antigen can be effectively reduced, and the linear range and accuracy of the novel coronavirus antigen detection can be improved, which is conducive to the prevention and control of the novel coronavirus.

[0043] In the embodiments of the present application, the minimum detection limit of the novel coronavirus antigen detection kit is 46.8 pg / mL. The novel coronavirus antigen detection kit provided in the present application has a low detection limit, and the novel coronavirus can be detected when it reaches 46.8 pg / mL, thereby facilitating the prevention and control of COVID-19.

[0044] In the embodiments of the present application, the linear range of the novel coronavirus antigen detection kit is 93.74 pg / mL-1200 pg / mL. The novel coronavirus antigen detection kit provided in the present application has a wide detection linear range and high accuracy, and can be applied to the detection of novel coronavirus in different scenarios.

[0045] In the present application, the capture antibody, the biotin-labeled detection antibody, and the SARS-CoV2 N protein standard can be solid or liquid, as long as they can stably exist in the detection kit.

[0046] In an embodiment of the present application, the capture antibody is solid, which can be dissolved in a suitable solution during the use of the detection kit. In another embodiment of the present application, the capture antibody is a solution containing anti-SARS-CoV2 N protein capture antibody, and the solute can be but is not limited to phosphate buffer. Further, the concentration of the anti-SARS-CoV2 N protein capture antibody in the solution containing the anti-SARS-CoV2 N protein capture antibody is 0.01 μg / mL-2 μg / mL. Specifically, the concentration of the anti-SARS-CoV2 N protein capture antibody in the solution containing the anti-SARS-CoV2 N protein capture antibody can be but is not limited to 0.01 μg / mL, 0.05 μg / mL, 0.1 μg / mL, 0.4 μg / mL, 0.8 μg / mL, 1 μg / mL, 1.5 μg / mL, or 2 μg / mL, etc.

[0047] In the present application, when detection is carried out by using the detection kit, the SARS-CoV2 N protein standard sample can be diluted to different concentrations to obtain a standard curve. In a specific embodiment, the SARS-CoV2 N protein standard sample is diluted to 46.87 pg / mL, 93.75 pg / mL, 187.5 pg / mL, 375 pg / mL, 750 pg / mL, 1500 pg / mL, 3000 pg / mL, 6000 pg / mL, and 12000 pg / mL.

[0048] In an embodiment of the present application, the detection antibody is in a solid state, which can be dissolved in a suitable solution during use of the detection kit. In another embodiment of the present application, the detection antibody is a solution of rabbit anti-SARS-CoV2 N protein antibody, wherein the solvent can be but is not limited to phosphate buffer. Further, the concentration of the rabbit anti-SARS-CoV2 N protein antibody in the solution of rabbit anti-SARS-CoV2 N protein antibody is 0.01 pg / mL to 2 pg / mL. Specifically, the concentration of the rabbit anti-SARS-CoV2 N protein antibody in the solution of rabbit anti-SARS-CoV2 N protein antibody can be but is not limited to 0.01 pg / mL, 0.05 pg / mL, 0.1 pg / mL, 0.4 pg / mL, 0.8 pg / mL, 1 pg / mL, 1.5 pg / mL, or 2 pg / mL, etc.

[0049] It can be understood that, if all reagents in the detection kit are in a solution state, the concentration of the reagents can be selected as needed, for example, a high-concentration reagent can be set, which can be diluted as needed during use of the detection kit; or the concentration of the reagent in the detection kit can be directly set to the concentration required for detection, thereby facilitating use.

[0050] In an embodiment of the present application, the detection kit further comprises a washing solution, which is a phosphate buffer solution (PBST) containing Tween. In the present application, the washing solution is used for the washing process during use of the detection kit, that is, during detection of the novel coronavirus antigen. In an embodiment of the present application, the concentration of the washing solution is 0.01% to 0.2%. Specifically, the concentration of the washing solution can be but is not limited to 0.01%, 0.05%, 0.08%, 0.1%, 0.12%, 0.15%, 0.18%, or 0.2%, etc. In a specific embodiment, 500 pL of Tween-20 can be added to 1 L of phosphate buffer solution to obtain the washing solution.

[0051] In the embodiments of the present application, the detection kit further comprises a blocking solution, and the blocking solution is a phosphate buffer solution containing bovine serum albumin and / or skimmed milk. The use of the above blocking solution can effectively block the uncoated binding sites in the detection hole which are not coated with the capture antibody, thereby improving the accuracy of the detection. In an embodiment of the present application, the blocking solution is a phosphate buffer solution containing bovine serum albumin, which is easy to store and use for a long time. Further, the concentration of the blocking solution is 0.5%-5%. Specifically, the concentration of the blocking solution can be, but is not limited to, 0.5%, 0.7%, 1%, 1.3%, 1.5%, 2%, 2.8%, 3%, 4% or 4.5%, etc. In a specific embodiment, 1 g of bovine serum albumin can be added to 100 mL of phosphate buffer solution to obtain the blocking solution.

[0052] In the embodiments of the present application, the enzyme-labeled avidin includes peroxidase-labeled avidin. Further, the enzyme-labeled avidin includes horseradish peroxidase-labeled avidin. Still further, the enzyme-labeled avidin includes horseradish peroxidase-labeled streptavidin (Streptavidin-HRP). In an embodiment of the present application, the enzyme-labeled avidin is in a liquid state, and the solvent can be, but is not limited to, a phosphate buffer solution. Further, the concentration of the liquid enzyme-labeled avidin is 0.05%-2%. Specifically, the concentration of the liquid enzyme-labeled avidin can be, but is not limited to, 0.05%, 0.07%, 0.1%, 0.2%, 0.5%, 1%, 1.2%, 1.5%, 1.8% or 2%, etc. In a specific embodiment, 1 mL of enzyme-labeled avidin can be dissolved in 200 mL of phosphate buffer solution to obtain the liquid enzyme-labeled avidin.

[0053] In the embodiments of the present application, the color developing solution includes a 3,3',5,5'-tetramethylbenzidine (TMB) solution. Under the action of peroxidase, the color of the TMB solution changes, so that the novel coronavirus in the sample to be detected can be qualitatively and quantitatively detected according to whether the color of the TMB solution changes and the absorbance of the solution before and after the color change.

[0054] In the embodiments of the present application, the termination solution includes a sulfuric acid solution. By adding the sulfuric acid solution, the activity of the enzyme molecules in the enzyme-labeled avidin can be destroyed, thereby stopping the reaction between the enzyme molecules and the color developing solution, and ensuring the accuracy of the detection results. In an embodiment of the present application, the concentration of the termination solution is 1M-4M. Specifically, the concentration of the termination solution can be, but is not limited to, 1M, 2M, 3M or 4M, etc.

[0055] In the embodiments of the present application, the detection kit further comprises an enzyme-labeled plate, and the enzyme-labeled plate is used to coat the capture antibody.

[0056] The novel coronavirus antigen detection kit provided in this application has a low detection limit, enabling detection of the novel coronavirus in the early stages of infection, thus effectively preventing and controlling COVID-19. At the same time, the detection kit can accurately detect the novel coronavirus, avoiding false positives and false negatives, and the detection is rapid, which is beneficial to its application.

[0057] This application also provides the application of the novel coronavirus antigen detection kit in any of the above embodiments in the detection of novel coronavirus for purposes other than disease diagnosis and / or treatment. The novel coronavirus antigen detection kit provided in this application can be used for the detection of novel coronavirus for purposes other than disease diagnosis and / or treatment, such as detecting novel coronavirus in the environment to determine whether the environment is safe, which is beneficial for the prevention and control of COVID-19.

[0058] This application also provides a method for detecting novel coronavirus antigens that is not intended for disease diagnosis and / or treatment, including detecting the novel coronavirus using a novel coronavirus antigen detection kit from any of the above embodiments.

[0059] Please see Figure 1 The flowchart of a method for detecting novel coronavirus antigen provided in one embodiment of this application includes:

[0060] S101: Disperse the capture antibody in phosphate buffer to obtain a capture antibody solution, and disperse the biotin-labeled detection antibody in phosphate buffer to obtain a detection antibody solution.

[0061] S102: Add the capture antibody solution to the detection well so that the capture antibody is coated in the detection well.

[0062] S103: Add the pretreated sample to the detection well for incubation, and then clean it.

[0063] S104: Add the detection antibody solution to the detection well and incubate, then wash.

[0064] S105: After adding enzyme-labeled avidin to the detection well, add the colorimetric solution to carry out the colorimetric reaction, and then add the stop solution to terminate the colorimetric reaction.

[0065] S106: Perform qualitative testing based on the color change of the detection orifice, and / or measure the OD of the detection orifice. 450 The value was compared with the standard curve to obtain the concentration of novel coronavirus antigen in the sample to be tested.

[0066] In the present application, by coating the capture antibody in the detection hole, the novel coronavirus in the sample to be tested is combined with the novel coronavirus in the sample to be tested during the incubation process, the biotin-labeled detection antibody is combined with the novel coronavirus in the sample to be tested, and the biotin-labeled detection antibody is combined with the enzyme-labeled avidin, thereby forming a complex; then color developing solution is added for color development, and after color development, stop solution is added to terminate the reaction, qualitative detection is carried out according to the color change, and quantitative detection can also be carried out according to the standard curve.

[0067] In the embodiments of the present application, the detection hole can be but is not limited to an enzyme-labeled plate.

[0068] In the embodiments of the present application, before the sample to be tested is added to the detection hole, the detection hole is also treated with a blocking solution, so that the sites in the detection hole that are not combined with the capture antibody are blocked.

[0069] In the embodiments of the present application, washing treatment is carried out using a washing solution.

[0070] In the present application, qualitative detection by color change of the detection hole includes: when the color developing solution is added, the reaction system changes from colorless to blue, and after the addition of the stop solution, it changes to yellow, indicating that the sample to be tested has the novel coronavirus; when the color developing solution is added, the color of the reaction system does not change, and after the addition of the stop solution, the color does not change, indicating that the sample to be tested does not have the novel coronavirus.

[0071] Please refer to Figure 2 The standard curve preparation flowchart of the novel coronavirus antigen detection method provided in an embodiment of the present application includes:

[0072] S201: Coating the capture antibody in a plurality of detection holes.

[0073] S202: Diluting the SARS-CoV2 N protein standard to form a plurality of SARS-CoV2 N protein solutions with different concentrations, adding the plurality of SARS-CoV2 N protein solutions with different concentrations to different detection holes respectively, and then washing.

[0074] S203: Adding the detection antibody solution to each detection hole for incubation, and then washing.

[0075] S204: Adding enzyme-labeled avidin to the detection hole, then adding color developing solution for color development reaction, and then adding stop solution to terminate the color development reaction.

[0076] S205: Measuring the OD 450 values of the plurality of detection holes, and obtaining a standard curve according to the SARS-CoV2 N protein solution concentration and the OD 450 values.

[0077] The method provided in the application has a wide linear range of the standard curve and a low detection limit, thereby facilitating detection.

[0078] In an embodiment of the application, the concentration ratio of the SARS-CoV2 N protein solution to the detection antibody is 4.68x10 -5 -1.2x10 -2 Using the above concentration ratio, the SARS-CoV2 N protein can be combined with the detection antibody efficiently, rapidly and specifically, the minimum detection limit is reduced, and the new coronavirus can be effectively detected at an early stage when the concentration of the new coronavirus is low.

[0079] In an embodiment of the application, the preparation of the standard curve comprises: adding 100 μl / well of the capture antibody to an enzyme-labeled plate, incubating at 4°C overnight, washing the plate 3 times with a washing solution (such as 0.05% PBST solution); adding 200 μl / well of a blocking solution (such as 1% BSA-PBS solution), incubating at 37°C for 1 hour, and washing the plate 3 times with the washing solution; diluting the SARS-CoV2 N protein standard to 46.87 pg / mL, 93.75 pg / mL, 187.5 pg / mL, 375 pg / mL, 750 pg / mL, 1500 pg / mL, 3000 pg / mL, 6000 pg / mL, and 12000 pg / mL, and adding 100 μl / well of each concentration to the enzyme-labeled plate, with three detection wells for each concentration, and incubating at 37°C for 2 hours; adding 100 μl / well of the biotin-labeled detection antibody, incubating at 37°C for 1 hour, and washing the plate 3 times with the washing solution; adding 100 μl / well of enzyme-labeled streptavidin (such as Streptavidin-HRP solution), incubating at 37°C for 30 minutes; adding 100 μl / well of a color developing solution (such as TMB solution), incubating in the dark for 20 minutes; adding 100 μl / well of a stop solution (such as sulfuric acid solution); and measuring the OD 450 value of each detection well; obtaining a standard curve according to the concentration of the SARS-CoV2 N protein solution and the OD 450 value, as shown in Figure 3 FIG. 1, wherein the horizontal coordinate of the standard curve is the concentration of the SARS-CoV2 N protein solution (pg / mL), the vertical coordinate is the OD 450 value, the standard curve is y=0.0002x+0.0139, R 2 =0.9986; the minimum detection limit is 46.87 pg / mL, and the linear range is 93.75 pg / mL-12000 pg / mL.

[0080] In an embodiment of the present application, the concentration of the washing solution is 0.01%-0.2%, the concentration of the sealing solution is 0.5%-5%, the concentration of the chromogenic solution is 0.01%-0.2%, and the concentration of the termination solution is 1M-4M. Specifically, the above-mentioned concentrations can be selected according to the detection needs.

[0081] The detection method of the novel coronavirus antigen provided by the present application is simple to operate, short in detection time, convenient in sampling, reduces the probability of false positives and false negatives, and improves the detection accuracy.

[0082] The above-described embodiments only express several embodiments of the present application, and the description is more specific and detailed, but it cannot be understood as limiting the scope of the patent of the present application. It should be noted that, for ordinary skilled persons in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are all within the protection scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims. SEQUENCE LISTING <110> SHENZHEN BINDE BIOLOGICAL TECHNOLOGY CO., LTD. <120> Novel coronavirus antigen detection kit and application, and novel coronavirus antigen detection method <160> 2 <170> SIPOSequenceListing 1.0 <210> 1 <211> 122 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 1 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Lys 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30 Ala Met His Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Lys Phe Thr Ile Ser Lys Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Ala Gln Asp Tyr Tyr Asp Asn Ser Gly Pro Ser Asp Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 2 <211> 108 <212> PRT <213> Artificial Sequence(Artificial Sequence) <400> 2 Val Ile Ile Thr Cys Arg Ala Pro Gln Thr Ile Ser Ser Trp Val Gly 1 5 10 15 Asp Lys Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Ile Ser Ala Cys 20 25 30 Lys Cys Leu Ala Trp Tyr Gln Gln Lys Pro Arg Lys Ala Pro Lys Leu 35 40 45 Leu Ile Tyr Lys Val Ser Ser Leu Glu Ser Gly Val Pro Ser Lys Phe 50 55 60 Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu 65 70 75 80 Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr 85 90 95 Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105

Claims

1. A test kit for a novel coronavirus antigen, characterized by, The capture antibody comprises an anti-SARS-CoV2 N protein capture antibody, a sequence of a heavy chain variable region of the anti-SARS-CoV2 N protein capture antibody is an amino acid sequence shown in SEQ ID NO: 1, and a sequence of a light chain variable region of the anti-SARS-CoV2 N protein capture antibody is an amino acid sequence shown in SEQ ID NO:

2.

2. The novel coronavirus antigen detection kit according to claim 1, characterized in that, The detection antibody is a rabbit anti-SARS-CoV2 N protein antibody with a product number of 40143-R040 from Beijing Yiqiao God Science and Technology Co., Ltd., and the SARS-CoV2 N protein standard product is an N protein standard product with a product number of B232004 from Taizhou Baiying Biological Technology Co., Ltd.

3. The novel coronavirus antigen detection kit according to claim 1, characterized in that, The minimum detection limit of the novel coronavirus antigen detection kit is 46.8 pg / mL, and the linear range is 93.74 pg / mL-1200 pg / mL.

4. The novel coronavirus antigen detection kit according to claim 1, wherein The enzyme-labeled affinity substance comprises a peroxidase-labeled affinity substance, the chromogenic solution comprises 3,3',5,5'-tetramethylbenzidine, and the termination solution comprises a sulfuric acid solution.

5. Application of the novel coronavirus antigen detection kit according to any one of claims 1-4 in detection of a novel coronavirus antigen without the purpose of disease diagnosis and / or treatment.

6. A method for detecting a novel coronavirus antigen for a purpose other than diagnosis and / or treatment of a disease, characterized by, The novel coronavirus antigen is detected by using the novel coronavirus antigen detection kit according to any one of claims 1-4.

7. The method of detecting a novel coronavirus antigen according to claim 6, wherein The kit comprises: The capture antibody is dispersed in a phosphate buffer solution to obtain a capture antibody solution, and the biotin-labeled detection antibody is dispersed in a phosphate buffer solution to obtain a detection antibody solution; The capture antibody solution is added to the detection hole to coat the capture antibody in the detection hole; The pretreated sample to be detected is added to the detection hole for incubation, and then washing is performed; The detection antibody solution is added to the detection hole for incubation, and then washing is performed; The enzyme-labeled affinity substance is added to the detection hole, and then the chromogenic solution is added for color development reaction, and then the termination solution is added to terminate the color development reaction; qualitative detection according to the color change of the detection well, and / or measuring the OD of the detection well 450 the concentration of the novel coronavirus antigen in the sample to be detected is obtained by comparing the value with a standard curve.

8. The method for detecting novel coronavirus antigen as described in claim 7, characterized in that, The preparation of the standard curve comprises: The capture antibody is coated in a plurality of detection holes; The SARS-CoV2 N protein standard product is diluted to form a plurality of SARS-CoV2 N protein solutions with different concentrations, and the plurality of SARS-CoV2 N protein solutions with different concentrations are added to different detection holes, respectively, and then washing is performed; The detection antibody solution is added to each detection hole for incubation, and then washing is performed; The enzyme-labeled affinity substance is added to the detection hole, and then the chromogenic solution is added for color development reaction, and then the termination solution is added to terminate the color development reaction; measuring the OD of a plurality of the detection wells 450 values, and obtaining the standard curve from the SARS-CoV2 N protein solution concentration and the OD 450 values.

9. The method for detecting novel coronavirus antigen as described in claim 8, characterized in that, The concentration ratio of the SARS-CoV2 N protein solution to the detection antibody is 4.68 x 10 -5 -1.2 x 10 -2 .

10. The method for detecting novel coronavirus antigen as described in claim 7, characterized in that, The concentration of the chromogenic solution is 0.01%-0.2%, and the concentration of the termination solution is 1M-4M.

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