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Kit for detecting novel coronavirus N protein and application thereof

A coronavirus and kit technology, applied in the field of kits for detecting the N protein of the new coronavirus, can solve the problems of insufficiently meeting the needs of epidemic prevention and control, and restricting the supply capacity of the rapid detection service of the new coronavirus, so as to reduce social panic. Emotion, fast detection speed, and the effect of accelerated screening

Inactive Publication Date: 2020-07-10
BEIJING ELCOTEQ BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On January 26, the State Drug Administration approved 4 new coronavirus detection products from 4 companies including BGI, but the currently approved new coronavirus nucleic acid detection reagents are all based on nucleic acid detection methods, and a single sample uses nucleic acid fluorescence. The PCR method takes 3 hours to give the test results, and the nucleic acid sequencing method takes 6 hours to give the test results. More importantly, due to the extremely high requirements of the laboratory environment for gene amplification, the detection throughput is called the biggest constraint factor, which is This greatly restricts the supply capacity of rapid detection services for the new coronavirus, which is not enough to fully meet the needs of epidemic prevention and control

Method used

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  • Kit for detecting novel coronavirus N protein and application thereof
  • Kit for detecting novel coronavirus N protein and application thereof
  • Kit for detecting novel coronavirus N protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 1. Preparation of novel coronavirus nucleocapsid (N) protein immunochromatographic detection kit

[0046] 1) Use fluorescent microspheres to label another strain of mouse anti-new coronavirus N protein monoclonal antibody

[0047] Take 0.5mL fluorescent microspheres, add 1mg carbodiimide (EDC), 1mg N-hydroxysuccinimide (NHS), stir at room temperature, 120r / min for 3h, then add 100μL of another strain of mouse anti-new coronavirus For virus N protein monoclonal antibody, stir for 1 hour at room temperature at a speed of 120 r / min, then add 10 mg of BSA blocking solution, and continue stirring for 1 hour at a speed of 120 r / min. Centrifuge at 12000 r / min for 20 min at 2-8 °C, and remove the supernatant. Finally, the solid precipitate obtained after centrifugation was redissolved to 1 mL with 0.2 M phosphate buffer (pH=7.4), and then 1 μL of Proclin300 was added and stored at 4°C until use.

[0048] 2) Using fluorescent microspheres to label rabbit IgG polyclonal antibod...

Embodiment 2

[0077] 1. Nasal / oropharyngeal swab sample testing

[0078] The types of samples taken included nasopharyngeal swabs and oropharyngeal swabs.

[0079] After the nasal / oropharyngeal swab samples are collected, they are pre-treated with the sample preservation solution (see Table 3 for the composition). Store in solution and stir. Squeeze the outside of the plastic hose several times with your fingers to fully soak the cotton swab with the sample preservation solution, then pull out the cotton swab, and the liquid that is wrung out (that is, the sample collected on the cotton swab is mixed into the sample preservation solution) is the sample to be tested . Then take 60 μL of the processed sample to be tested and drop it into the sample hole of the test card, let it stand for 15 minutes, and then insert it into the fluorescence immunochromatography analyzer for detection, the instrument automatically calculates the T / C value of the sample, and passes the normal range Judge nega...

Embodiment 3

[0087] Example 3 Serum / Plasma / Whole Blood Sample Detection

[0088] The types of samples taken include serum / plasma / whole blood.

[0089] After the serum / plasma / whole blood sample is collected, no sample pretreatment is required, and the sample to be tested is directly added dropwise to the sample inlet of the test card and allowed to stand for 15 minutes, and then inserted into the fluorescence immunochromatography analyzer for detection, the instrument automatically calculates the sample T / C value, judge negative or positive by comparing with normal value range. In addition, when two fluorescent bands appear under ultraviolet light irradiation, it is positive. The clinical test results are shown in Table 5 below.

[0090] table 5

[0091]

[0092]

[0093] As can be seen from the clinical test data in Table 5, all 20 negative samples were detected as negative, which was consistent with the clinical diagnosis; all 20 positive samples were tested positive, with a posi...

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Abstract

The invention relates to a kit for detecting novel coronavirus N protein and an application of the kit. The kit comprises (1) one antibody capable of being bound to the novel coronavirus N protein, and (2) the other antibody marked by a marker and capable of being bound to the novel coronavirus N protein when the novel coronavirus N protein can be bound to (1) to limit the antibody. The using method of the kit comprises the step of determining whether a patient suffers from the novel coronavirus pneumonia or not by detecting the level of the novel coronavirus N protein in an individual sample.The kit for detecting the novel coronavirus N protein provided by the invention can be used for detecting the coronavirus N protein in nasopharynx swabs, oropharynx swabs, alveolar lavage fluids, serum, plasma, whole blood, sputum, oral swabs or saliva samples so as to be used for diagnosing novel coronavirus pneumonia.

Description

technical field [0001] The invention belongs to the field of in vitro diagnosis, and relates to a kit for detecting the N protein of novel coronavirus and its application 。 Background technique [0002] The 2019 novel coronavirus (SARS-CoV-2) is a new strain of coronavirus that has never been found in humans before. Common signs of human infection with coronavirus include respiratory symptoms, fever, cough, shortness of breath, and dyspnea. In more severe cases, infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and even death. On January 26, the State Drug Administration approved 4 new coronavirus detection products from 4 companies including BGI, but the currently approved new coronavirus nucleic acid detection reagents are all based on nucleic acid detection methods, and a single sample uses nucleic acid fluorescence. The PCR method takes 3 hours to give the test results, and the nucleic acid sequencing method takes 6 hours to give the t...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/165G01N2469/10
Inventor 林斯
Owner BEIJING ELCOTEQ BIO TECH
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