CN112553077A - Novel coronavirus lysate and application thereof - Google Patents
Novel coronavirus lysate and application thereof Download PDFInfo
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- CN112553077A CN112553077A CN202011463511.9A CN202011463511A CN112553077A CN 112553077 A CN112553077 A CN 112553077A CN 202011463511 A CN202011463511 A CN 202011463511A CN 112553077 A CN112553077 A CN 112553077A
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- novel coronavirus
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- lysate
- antigen
- colloidal gold
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- C12N1/06—Lysis of microorganisms
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Abstract
The invention provides a novel coronavirus 2019-nCoV lysate and application thereof, wherein the lysate comprises the following components: Tris-HCl 1.6-2.4mol/L, SDS mass percent is 0.8-1.2%, polyvinylpyrrolidone mass percent is 0.4-0.6%, and pH8.0-9.0; the novel coronavirus 2019-nCoV lysate can be used for fully cracking the novel coronavirus 2019-nCoV, so that the nucleocapsid protein (N protein) of the novel coronavirus 2019-nCoV is fully exposed, and the sensitivity and specificity for detecting the novel coronavirus 2019-nCoV antigen are improved.
Description
Technical Field
The invention relates to the field of virus detection, in particular to a novel coronavirus lysate and application thereof.
Background
2019 novel coronavirus, which is formally named 2019-nCoV by the world health organization in 1 month and 12 days 2020. Coronaviruses are a large family of viruses known to cause the common cold and more serious diseases such as Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). The novel coronavirus is a new strain of coronavirus that has not been previously discovered in humans.
The common signs of human infection with the novel coronavirus include respiratory symptoms, fever, cough, shortness of breath, dyspnea and the like. In more severe cases, the infection can lead to pneumonia, severe acute respiratory syndrome, renal failure, and even death. Many symptoms are treatable and therefore need to be treated according to the clinical condition of the patient. In addition, the adjuvant care of infected persons can be very effective.
At present, test paper strips are adopted to detect novel coronavirus infection, and novel coronavirus antibody IgM/IgG antibody is generally detected. The techniques for directly detecting the novel coronavirus antigen are few, because the novel coronavirus load in the collected sample is low and the target protein to be detected in the particles, such as the nucleocapsid protein (N protein), is not fully exposed, which results in the defects of poor sensitivity and specificity during detection.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide a novel coronavirus 2019-nCoV lysate and application thereof.
The invention provides a novel coronavirus 2019-nCoV lysate, which comprises the following components: Tris-HCl 1.6-2.4mol/L, SDS (sodium dodecyl sulfate) 0.8-1.2 wt%, polyvinyl pyrrolidone 0.4-0.6 wt%, and pH 8.0-9.0.
The novel coronavirus 2019-nCoV lysate comprises the following components: Tris-HCl 2mol/L, SDS wt% and polyvinyl pyrrolidone 0.5 wt%, and has pH of 8.0-9.0.
The invention provides application of the novel coronavirus 2019-nCoV lysate in preparation of a novel coronavirus antigen or a product for detecting the novel coronavirus 2019-nCoV antigen.
The novel coronavirus 2019-nCoV antigen is novel coronavirus 2019-nCoV nucleocapsid protein.
The invention provides a kit for detecting a novel coronavirus 2019-nCoV antigen, which comprises a novel coronavirus 2019-nCoV lysate as set forth in claim 1 or claim 2.
The kit for detecting the novel coronavirus 2019-nCoV antigen comprises an immunochromatography test strip for detecting the novel coronavirus 2019-nCoV antigen.
The invention provides a method for preparing a novel coronavirus 2019-nCoV antigen, which comprises the following steps:
obtaining a sample containing the novel coronavirus 2019-nCoV;
adding the sample to a lysate of the novel coronavirus 2019-nCoV of claim 1 or 2.
According to the method for preparing the novel coronavirus 2019-nCoV antigen, the cracking condition is that the treatment is carried out for 1-3 minutes; optionally for 2 minutes.
The technical scheme of the invention has the following advantages:
1. the invention provides a novel coronavirus 2019-nCoV lysate which comprises the following components: Tris-HCl 1.6-2.4mol/L, SDS mass percent is 0.8-1.2%, polyvinylpyrrolidone mass percent is 0.4-0.6%, and pH8.0-9.0; the novel coronavirus 2019-nCoV lysate can fully lyse the novel coronavirus 2019-nCoV, so that the nucleocapsid protein (N protein) of the novel coronavirus 2019-nCoV is fully exposed for detection, and if an immunochromatography test strip for detecting the novel coronavirus 2019-nCoV antigen is adopted, the exposed nucleocapsid protein (N protein) can be combined with the novel coronavirus 2019-nCoV antibody marked by the colloidal gold on the colloidal gold adsorption pad and the novel coronavirus 2019-nCoV antibody coated on a detection line, so that the sensitivity and specificity for detecting the novel coronavirus 2019-nCoV antigen are improved.
2. The invention provides a method for preparing a novel coronavirus 2019-nCoV antigen, which comprises the following steps: obtaining a sample containing the novel coronavirus 2019-nCoV; adding said sample to said novel coronavirus 2019-nCoV lysate; the method can be used for preparing the novel coronavirus 2019-nCoV antigen.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
Fig. 1 is a schematic structural diagram of a novel coronavirus antigen detection test strip according to an embodiment of the invention.
Reference numerals:
1-substrate, 2-loading pad, 3-colloidal gold adsorption pad, 4-antibody bearing membrane, 5-water absorption pad, 6-protective membrane, 7-blood filtering membrane, T-detection line T, C-quality control line C.
Detailed Description
The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
When the reagent is used for detection, for example, an immunochromatographic test strip for detecting the novel coronavirus 2019-nCoV antigen is adopted, the exposed nucleocapsid protein (N protein) can be combined with the novel coronavirus 2019-nCoV antibody marked by the colloidal gold on the colloidal gold adsorption pad and the novel coronavirus 2019-nCoV antibody coated on a detection line.
Example 1 novel coronavirus 2019-nCoV lysate
The embodiment provides a novel coronavirus 2019-nCoV lysate, which has the following formula:
1.2 percent of Tris-HCl 1.6mol/L, SDS, 0.4 percent of polyvinylpyrrolidone and the balance of water, and the pH value is 8.0-9.0.
The novel coronavirus 2019-nCoV lysate is prepared according to a conventional method.
Example 2 novel coronavirus 2019-nCoV lysate
The embodiment provides a novel coronavirus 2019-nCoV lysate, which has the following formula:
Tris-HCl2.4mol/L, SDS accounts for 0.8 percent by mass, polyvinylpyrrolidone accounts for 0.6 percent by mass, and the balance is water with the pH value of 8.0-9.0.
The novel coronavirus 2019-nCoV lysate is prepared according to a conventional method.
Example 3 novel coronavirus 2019-nCoV lysate
The embodiment provides a novel coronavirus 2019-nCoV lysate, which has the following formula:
Tris-HCl 2mol/L, SDS wt% and polyvinyl pyrrolidone 0.5 wt%, except water at pH 8.0-9.0.
The novel coronavirus 2019-nCoV lysate is prepared according to a conventional method.
Example 4 kit for detecting novel coronavirus 2019-nCoV antigen
The embodiment provides a kit for detecting a novel coronavirus 2019-nCoV antigen, which comprises the lysate of embodiment 1, embodiment 2 or embodiment 3, wherein the lysate of embodiment 3 is selected in the embodiment.
Further, the kit also comprises an immunochromatographic test strip for detecting the novel coronavirus 2019-nCoV antigen, as shown in figure 1, a sample loading pad 2, a colloidal gold adsorption pad 3, a hemofiltration membrane 7, an antibody bearing membrane 4 and a water absorption pad 5 are sequentially overlapped and lapped on a substrate 1; the upper surfaces of the sample loading pad 2 and the water absorption pad 5 are provided with protective films 6;
the antibody bearing film 4 is provided with a detection line T and a quality control line C which are spaced, the detection line T is close to the colloidal gold adsorption pad, and the quality control line C is close to the water absorption pad;
the coating of the colloidal gold adsorption pad 3 is coated with a colloidal gold-labeled novel coronavirus monoclonal antibody B and a colloidal gold-labeled mouse IgG antibody;
a novel coronavirus monoclonal antibody A is coated on the detection line T;
the quality control line C is coated with an anti-mouse IgG polyclonal antibody.
The preparation method of the immunochromatographic test strip for detecting the novel coronavirus 2019-nCoV antigen comprises the following steps:
1. preparing an antibody bearing membrane:
(1) selecting a nitrocellulose membrane with the aperture of 3-10 mu m, and cutting the membrane into specifications with the width of 2.0cm and the length of 30.5cm according to requirements for later use.
(2) Preparing a novel coronavirus monoclonal antibody A for coating a detection line by using a Tris-HCl buffer solution with the pH value of 0.1M and the pH value of 8.0, wherein the antibody concentration is 1.2-1.5 mg/ml, and in the embodiment, the antibody concentration is 1.3 mg/ml; sodium chloride buffer solution with the mass percentage content of 0.85% is used for preparing the anti-mouse IgG polyclonal antibody for quality control line coating, so that the antibody concentration is (1.5-1.8) mg/ml, and in the embodiment, the antibody concentration is 1.6 mg/ml.
(3) Selecting an antibody coating surface of a nitrocellulose membrane and marking, coating the detection line to be coated and the antibody solution of the quality control line on the membrane in parallel and uniformly, and controlling the distance between the detection line and the quality control line to be C-IgG: 4.0mm, and drying the nitrocellulose membrane at the constant temperature of 2-30 ℃ for later use.
(4) Preparing a sealing treatment soaking solution, and adding purified water with actual production capacity into a liquid preparation tank; respectively weighing buffer solution, sugar, blocking protein and preservative, directly adding the above components into a liquid preparation tank, stirring until the components are completely dissolved, adding purified water to a desired volume, and fully and uniformly stirring for at least 10 minutes for later use; the prepared sealing treatment soaking solution contains 0.1Mol of buffer solution, 0.5 mass percent of sugar, 1 mass percent of sealing protein and 0.05 mass percent of preservative, wherein the buffer solution is phosphate buffer solution, the sugar is cane sugar, the preservative is thimerosal, and the sealing protein is bovine serum albumin.
(5) Placing the membranes coated with the detection lines and the quality control lines in a treatment tank, adding the prepared sealing treatment soaking liquid of the step (4), ensuring that each membrane is completely immersed in the sealing treatment soaking liquid of the step (4) for 30 minutes, ensuring that the membranes do not move and overlap, taking out the membranes from the treatment tank after 30 minutes, and pouring the sealing treatment soaking liquid of the step (4); and (3) placing the membrane on gauze by using tweezers, and airing to be slightly dry to obtain the antibody bearing membrane.
(6) Pasting a board and drying: tearing off white paper in the middle of a cutting line on double-sided adhesive of a rubber plate, putting an antibody bearing film subjected to sealing treatment right at the central blank position of the rubber plate by using tweezers, enabling the right side of the rubber plate to be flush with the right side of the film, avoiding errors in the production process, ensuring that the color development position is relatively accurate, pasting one end of all top quality control lines when pasting the plate, smearing the film surface through the double-sided adhesive paper after the film is pasted on the rubber plate, avoiding bubbles, controlling the temperature in a control room to be 18-28 ℃ and the relative humidity to be less than or equal to 40%, and ensuring that air can circulate in a drying room and air of a dehumidifier can not directly blow on the film surface. The drying time is more than or equal to 4 hours and is reserved.
2. Preparation of colloidal gold adsorption pad
(1) Preparing a colloidal gold complex solution: adding an actual production amount of purified water to the liquor preparation tank; trehalose, bovine serum albumin, trisodium citrate, polyethylene glycol and NaN were weighed using an electronic analytical balance3Directly adding the colloidal gold complex solution into a liquid preparation tank for stirring until the colloidal gold complex solution is completely dissolved, adding purified water to a constant volume to a required volume, fully and uniformly stirring, wherein the stirring time is more than 30 minutes, and the obtained colloidal gold complex solution contains 5 mass percent of trehalose, 2 mass percent of bovine serum albumin, 0.5 mass percent of trisodium citrate, 0.05 mass percent of polyethylene glycol and 0.05 mass percent of NaN3。
(2) Measuring colloidal gold with required labeling amount by using a measuring cylinder, adjusting according to pH7.0-7.3, namely adding 0.2mol/L potassium carbonate solution according to the volume percentage content of 0.50%, stirring for 15 minutes on a magnetic stirrer, then taking a novel coronavirus monoclonal antibody B, diluting the novel coronavirus monoclonal antibody B by using double distilled water, labeling the colloidal gold according to 10-12 mu g/ml (namely, after adding the colloidal gold into the novel coronavirus monoclonal antibody B dilution, the final concentration of the novel coronavirus monoclonal antibody B is 10-12 mu g/ml, in the embodiment, the final concentration is 11 mu g/ml), adding the novel coronavirus monoclonal antibody B into the colloidal gold, stirring for 30 minutes on the magnetic stirrer, adding 0.5 per thousand stabilizer polyethylene glycol according to the volume percentage content, stirring for 30 minutes, then centrifuging, collecting precipitates, re-dissolving the colloidal gold compound solution according to the volume percentage of 3 percent, and stirring the mixture on a magnetic stirrer until the mixture is uniformly mixed to obtain the colloidal gold-novel coronavirus monoclonal antibody B conjugate compound solution with the volume percentage of 3 percent for later use.
(3) Measuring colloidal gold with a required labeling amount by using a measuring cylinder, adjusting according to pH7.0-7.3, namely adding 0.2mol/L potassium carbonate solution according to the volume percentage content of 0.50%, stirring for 15 minutes on a magnetic stirrer, taking a mouse IgG antibody, diluting the mouse IgG antibody by using double distilled water, labeling the colloidal gold according to the volume percentage content of 3-5 mu g/ml (namely, after adding the colloidal gold into the mouse IgG antibody diluent, the final concentration of the mouse IgG antibody is 3-5 mu g/ml, in the embodiment, the final concentration is 4 mu g/ml), adding the mouse IgG antibody into the colloidal gold, stirring for 30 minutes on the magnetic stirrer, adding 0.5 thousandth of stabilizer polyethylene glycol according to the volume percentage content, stirring for 30 minutes, centrifuging, collecting precipitates, re-dissolving the colloidal gold re-solution according to the volume percentage content of 3%, stirring on the magnetic stirrer until the mixture is uniform, obtaining the colloidal gold-mouse IgG antibody re-combination solution with the volume percentage content of 3%, and (5) standby.
(4) Taking the colloidal gold-novel coronavirus monoclonal antibody B conjugate compound solution with the volume percentage content of 3% and the colloidal gold-mouse IgG antibody conjugate compound solution with the volume percentage content of 3% in the above (2) and (3), re-dissolving the colloidal gold re-solution with the volume percentage content of 50%, uniformly mixing the re-dissolved colloidal gold-novel coronavirus monoclonal antibody B conjugate compound solution and the colloidal gold-mouse IgG antibody conjugate compound solution on a magnetic stirrer according to the volume percentage content of 50%, and uniformly mixing the re-dissolved colloidal gold-solution and the2Pouring the colloidal gold onto a prepared colloidal gold adsorption pad, placing the colloidal gold adsorption pad in a drying chamber to be dried for more than or equal to 4 hours, controlling the temperature of the drying chamber to be 18-28 ℃, controlling the relative humidity to be less than or equal to 40%, ensuring smooth air and preventing airflow from directly blowing on the colloidal gold adsorption pad, placing the dried colloidal gold adsorption pad into an aluminum foil bag filled with a drying agent, sealing and storing, and making a labeled colloidal gold adsorption pad for later use.
Note: the colloidal gold is cast because the cast colloidal gold can be freely cut in width, thereby being convenient for adjusting the color development of the product.
3. Assembling and cutting:
(1) taking a semi-finished product of a transparent substrate 1 adhered with an antibody bearing film, cutting a blood filtering film 7 into 0.5cm in width, adhering the blood filtering film to the transparent substrate close to one side of a detection line T by 0.5cm in width, keeping the blood filtering film to be in lap joint with the antibody bearing film 4 by about 1mm, cutting a test strip colloidal gold adsorption pad 3 into 0.5cm in width, adhering the strip to the transparent substrate 1 of the blood filtering film 7 far away from the detection line T, keeping the test strip colloidal gold adsorption pad to be in lap joint with the blood filtering film 7 by about 1mm, compounding a test strip water absorption pad 5 on the transparent substrate close to one side of a quality control line C and in lap joint with the antibody bearing film 4 by about 1mm, compounding a test strip upper sample pad 2 on one end of the test strip colloidal gold adsorption pad 3 far away from the blood filtering film 7 and in lap joint with the test strip colloidal gold adsorption pad by about 1mm, and marking for;
(2) and cutting the assembled substrate into strip test paper for later use, and respectively attaching protective films 6 to the sample loading pad 2 and the water absorption pad 5.
Example 5 detection method of novel coronavirus 2019-nCoV antigen
The embodiment provides a method for detecting a novel coronavirus 2019-nCoV antigen, which comprises the following steps: before testing, the test strips, samples, and lysates need to be kept at room temperature (15-30 ℃) and used as soon as possible.
1. The sample collection swab was inserted into an extraction flask containing 250ul of a lysate (a mixed aqueous solution containing 1% by mass of Tris-HCl 2mol/L, SDS and 0.5% by mass of polyvinylpyrrolidone, pH8-9), and the swab was repeatedly rotated at least 1-5 times (4 times selected in this example) for 2 minutes.
2. The swab is squeezed at the tube wall, causing the liquid to be continuously expressed. The sample is removed and the swab discarded according to the infected item disposal method.
3. The extraction flask was capped with a distributor head. (if the treated sample is used within 60 minutes, the test result of the kit is not affected)
4. The test strip was removed from the sealed foil pouch and placed on a clean flat surface. 2 drops of the treated sample were added vertically onto the loading pad of the test strip.
5. Wait for 15 minutes, interpret and record the test results. After 20 minutes the results were not valid.
And (4) interpretation of results: when a positive novel coronavirus antigen sample is detected, a purple red strip is displayed at the detection line T, and a purple red strip is displayed at the quality control line C. Negative specimens showed only a purple-red band at the control line C.
Comparative example 1
The embodiment provides a novel coronavirus 2019-nCoV lysate, which has the following formula:
Tris-HCl 2mol/L, SDS mass percent is 1%, the rest is water, pH8.0-9.0.
Comparative example 2
The embodiment provides a novel coronavirus 2019-nCoV lysate, which has the following formula:
Tris-HCl 2mol/L, polyvinylpyrrolidone 0.5 wt%, and water in balance, and the pH value is 8.0-9.0.
Comparative example 3
The embodiment provides a novel coronavirus 2019-nCoV lysate, which has the following formula:
Tris-HCl 4mol/L, SDS wt% 1.5 wt%, polyvinyl pyrrolidone 1.5 wt%, and water in balance, and has pH of 8.0-9.0.
Experimental example 1
1. Experimental reagent
The in vitro diagnostic reagent reference is used for detection according to the example 5 (since the reference is used as a sample to be detected, the step 1-2 of the example 5 is directly replaced by the reference according to the following using method and requirements, the lysate of the example 3 is used as a diluent during dilution, and detection is carried out after dilution), the in vitro diagnostic reagent reference is used: national reference of reagents for detecting antigens of new coronavirus (national institute of food and drug testing):
[ batch No. ] 370095-202001
[ PROPERTIES ] A liquid
The reference material is prepared by diluting a common buffer solution containing phosphate buffer solution, serum albumin (about 1%), trehalose (or sucrose, lactose, 5%) and gelatin (or gelatin hydrolysate, dextran, 1%). Is suitable for the quality control and evaluation of novel coronavirus antigen detection reagents (including but not limited to colloidal gold method, immunofluorescence method, chemiluminescence method and the like).
[ COMPOSITION AND SPECIFICATION ]
[ METHOD OF USE AND DEMAND ] OF USE
1. The using method comprises the following steps:
1) positive reference products P1-P8: diluting the reagent diluent or pure water by 1: 10 and then detecting;
2) negative reference products N1-N20: directly using the original times for detection;
3) minimum detection limit reference S: diluting the reagent diluent or pure water by 1: 50, 1: 100, 1: 200, 1: 400, 1: 800 and 1: 1600, marking as S1-S6, and respectively detecting the 6 concentrations;
4) repetitive reference R: the concentrations were measured 10 times, respectively, using a 1: 10 and a 1: 100 dilution of each reagent diluent or pure water, and labeled as R1 and R2.
2. When the reference product is used, the following requirements are met:
1) positive compliance rate: the results of P1-P8 are positive, and the coincidence rate is 8/8;
2) negative coincidence rate: N1-N20 are all negative, and the coincidence rate is 20/20;
3) the lowest detection limit is: the S1-S4 are all positive, and S5 and S6 are not required;
4) repeatability: the 10 detection results of R1 and R2 are positive, and the color development degree is uniform and has no difference; or a CV of no greater than 20.0% for 10 test results (if applicable, if required by relevant industry standards executable standards).
Screw mouth plastic freezing tube
[ STORAGE ] the long-term storage should be at-80 ℃ or below.
[ notes ] to provide a novel therapeutic agent
1. The reference should be stored at-80 deg.C and below. The split charging is carried out before the use is recommended, and repeated freeze thawing for more than 3 times is avoided;
2. raw materials used by the reference product are subjected to inactivation treatment, but the inactivation efficiency of the reference product is not fully verified, the components are still treated as potential infection sources in the using process, and the operation is executed according to laboratory safety management regulations;
3. the reference substances N1-N20 and P5-P7 are inactivated by heat at 56 ℃ or 65 ℃, and P1-P4, S and R are inactivated by beta-propiolactone.
2. Physical properties:
2.1 appearance Properties
The appearance is smooth, the plastic shell is firmly matched, and the assembly of the test paper in the shell is smooth and regular.
2.2 film strip Width
The film strip should be wider than 2.5 mm.
2.3 speed of travel
The liquid moving speed is not lower than 10 mm/min.
3. The lowest detection limit is:
the lysate test results of example 3 were used: the detection is carried out by using the lowest detection limit reference substance of enterprises or countries, S1, S2, S3 and S4 are all positive reactions, S5 is a positive reaction, and S6 is a negative reaction.
4. Specificity:
4.1 lysate test result of example 3:
negative reference product compliance rate: the detection is carried out by using negative reference substances of enterprises or countries, N1-N20 are all negative, and the coincidence rate (-/-) is 20/20.
Positive reference compliance rate: the detection is carried out by using enterprise or national positive reference substances, the P1-P8 are all positive, and the compliance rate (+/+) is 8/8.
4.2 detection results with the lysate of comparative example 1:
negative reference product compliance rate: the detection is carried out by using negative reference substances of enterprises or countries, N1-N20 are all negative, and the coincidence rate (-/-) is 20/20.
Positive reference compliance rate: the test is carried out by using an enterprise or national positive reference substance, and the coincidence rate (/ +) is 5/8.
4.3 lysate test result of comparative example 2:
negative reference product compliance rate: the detection is carried out by using negative reference substances of enterprises or countries, N1-N20 are all negative, and the coincidence rate (-/-) is 20/20.
Positive reference compliance rate: the test is carried out by using an enterprise or national positive reference substance, and the coincidence rate (/ +) is 3/8.
4.3 lysate test result of comparative example 3:
negative reference product compliance rate: the detection is carried out by using negative reference substances of enterprises or countries, N1-N20 are all negative, and the coincidence rate (-/-) is 20/20.
Positive reference compliance rate: the test is carried out by using an enterprise or national positive reference substance, and the coincidence rate (/ +) is 6/8.
5. Repeatability:
the lysate test results of example 3 were used: the repeated detection is respectively carried out for 10 times by using enterprise or national repetitive reference products (R1 and R2), the results are positive, the reaction results are consistent, and the color development degree is uniform.
6. And (3) testing the stability:
and (3) placing the test paper at 37 ℃ for 20 days, or placing the test paper at a room temperature sealed and dark dry place for 12 months, and detecting the appearance shape, the width of the membrane strip, the migration speed, the minimum detection limit, the reference product compliance rate and the repeatability of the test paper which meet the regulation requirements of 2-2.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (8)
1. A novel coronavirus 2019-nCoV lysate, which is characterized by comprising the following components: Tris-HCl 1.6-2.4mol/L, SDS mass percent is 0.8-1.2%, polyvinylpyrrolidone 0.4-0.6%, pH8.0-9.0.
2. The novel coronavirus 2019-nCoV lysate according to claim 1, which comprises the following components: Tris-HCl 2mol/L, SDS wt% and polyvinyl pyrrolidone 0.5 wt%, and has pH of 8.0-9.0.
3. Use of the novel coronavirus 2019-nCoV lysate according to claim 1 or 2 for the preparation of a novel coronavirus antigen or for the preparation of a product for the detection of a novel coronavirus 2019-nCoV antigen.
4. Use according to claim 3, characterized in that the novel coronavirus 2019-nCoV antigen is a novel coronavirus 2019-nCoV nucleocapsid protein.
5. A kit for detecting a novel coronavirus 2019-nCoV antigen, which comprises the novel coronavirus 2019-nCoV lysate of claim 1 or 2.
6. The kit for detecting the novel coronavirus 2019-nCoV antigen as claimed in claim 5, which is characterized by comprising an immunochromatographic test strip for detecting the novel coronavirus 2019-nCoV antigen.
7. A method for preparing a novel coronavirus 2019-nCoV antigen, which comprises the following steps:
obtaining a sample containing the novel coronavirus 2019-nCoV;
adding the sample to a lysate of the novel coronavirus 2019-nCoV of claim 1 or 2.
8. The method for preparing the novel coronavirus 2019-nCoV antigen as claimed in claim 7, wherein the lysis conditions are treatment for 1-3 minutes; optionally for 2 minutes.
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