Novel coronavirus earlier-stage screening method

A coronavirus, a new type of technology, applied in the field of immune detection, can solve problems such as increased detection value, inability to effectively distinguish patients, and inability to meet the needs of patients screened out, so as to achieve accurate detection results

Active Publication Date: 2020-07-03
GUANGZHOU KANGRUN BIOTECHNOLOGY CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection rate of nucleic acid detection reagents for new coronary pneumonia is low, and the sensitivity is only 30-50%, which cannot meet the needs of screening out all patients
[0005] The detection of total antibodies improves the sensitivity but cannot effectively distinguish which stage the patient is in and whether they are still infectious
The colloidal gold method is simple and fast, but the accuracy is not enough
The sensitivity and specificity of IgM and IgG antibodies measured by chemiluminescence method are both high, but the window period of the two is relatively long. IgM should be detected 3-5 days after the onset of the disease, and IgG can only be diagnosed after the detection value increases by more than 4 times during the recovery period.
At present, there is no report on the use of IgA in the screening and detection of new coronary pneumonia, and there is no report on the successful development of an IgA detection kit

Method used

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  • Novel coronavirus earlier-stage screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A kit for screening novel coronavirus infection for detection of SARS-CoV-2 IgA antibody. Especially for early screening of SARS-CoV-2 IgA antibodies.

[0037] The kit for screening novel coronavirus infection includes:

[0038]1. SARS-CoV-2 magnetic bead coating (R1): containing the magnetic bead coating coated with SARS-CoV-2 recombinant antigen Tris buffer, pH 7.1-7.4, containing 0.05 -0.2% Tween 20 (v / v), 2-8% bovine serum albumin (w / v), 0.25-1% Proclin300 (v / v); magnetic beads are used as the solid phase carrier for the reaction, and the antigen is used to bind the target antibody in the assay.

[0039] 2. Anti-human IgA antibody acridinium ester label (R2): 2-(N-morpholine) ethanesulfonic acid buffer for acridine ester-labeled mouse anti-human IgA monoclonal antibody, containing 0.05-0.2% Tween 20 (v / v), 2-8% bovine serum albumin (w / v), 0.25-1% Proclin300 (v / v). Combined with the antigen-antibody complex, under alkaline conditions, the acridinium ester can emi...

Embodiment 2

[0048] A kit for detecting SARS-CoV-2 IgA antibody for screening new coronavirus infection, other features are the same as in Example 1, the difference is that it also has the following features: SARS-CoV-2 recombinant antigen is suspended in suspension The Spike protein (S protein) expressed in the cultured human embryonic kidney cell line HEK293F, referred to as S protein for short, was truncated from the amino acid sequence of Gln321-Ser591 in the RBD domain for protein expression. The protein expression process is as follows: the SARS-CoV-2 S protein RBD gene is linked to human IgG1 Fc through a TEV restriction site sequence at the C-terminus, and then cloned into the mammalian expression vector pTT5 and the Kozak sequence is added to the translation initiation region , and then transfected into suspension culture cells HEK293F, using interferon α-1 as the signal peptide to make HEK293F cells secrete RBD-TEV-Fc fusion protein, and then use Protein A column and nickel column...

Embodiment 3

[0065] A kit for screening SARS-CoV-2 IgA antibodies for screening novel coronavirus infection, especially for screening SARS-CoV-2 IgA antibodies at an early stage.

[0066] This kit for screening for novel coronavirus infection contains:

[0067] 1. SARS-CoV-2 magnetic bead coating (R1): a magnetic bead coating containing SARS-CoV-2 recombinant antigen Tris buffer, pH 7.2, containing 0.1% NaCl Warm 20 (v / v), 5% bovine serum albumin (w / v), 0.5% Proclin300 (v / v); magnetic beads are used as the solid phase carrier for the reaction, and the antigen is used to bind the target antibody in the analyte.

[0068] 2. Anti-human IgA antibody acridinium ester label (R2): 2-(N-morpholine) ethanesulfonic acid buffer for acridine ester-labeled mouse anti-human IgA monoclonal antibody, containing 0.1% Tween 20 (v / v), 5% bovine serum albumin (w / v), 0.5% Proclin300 (v / v). Combined with the antigen-antibody complex, under alkaline conditions, the acridinium ester can emit light after being ...

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Abstract

A novel coronavirus earlier-stage screening method disclosed by the invention can be used for detecting the novel coronavirus, and is particularly used for screening the SARS-CoV-2IgA antibody in theearly stage. A kit comprises an SARS-CoV-2 magnetic bead coating material used as R1, containing a magnetic bead coating material trihydroxymethyl aminomethane buffer solution coated by an SARS-CoV-2recombinant antigen, wherein the pH value is 7.1-7.4; and an acridinium ester marker of the anti-human IgA antibody as R2, wherein the acridinium ester marker is a 2-(N-morpholine) ethanesulfonic acidbuffer solution containing an acridinium ester labeled mouse anti-human IgA monoclonal antibody. The kit and the SARS-CoV-2IgA antibody immunoassay reagent disclosed by the invention can be used forsupplementing and diagnosing new coronal pneumonia at an earlier stage, and have the characteristic of accurate detection result.

Description

technical field [0001] The present invention relates to the technical field of immunoassay, in particular to an earlier screening method, reagent and kit for SARS-CoV-2 IgA antibodies of novel coronavirus-infected pneumonia (COVID-19). Background technique [0002] As an acute respiratory infectious disease, novel coronavirus-infected pneumonia (COVID-19) has been included in the Class B infectious disease stipulated in the "Law of the People's Republic of China on the Prevention and Control of Infectious Diseases", and is managed as a Class A infectious disease. COVID-19 usually develops 3-7 days after infection with SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2). Symptoms include fever, dry cough, and fatigue, and may also be accompanied by nasal congestion, runny nose, sore throat, myalgia, and diarrhea. . Mild patients have no symptoms of pneumonia, severe patients often develop dyspnea and / or hypoxemia one week after the onset, and severe patients can rap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/569G01N33/558G01N33/543G01N21/76
CPCG01N21/76G01N33/54326G01N33/558G01N33/56983G01N33/577G01N33/587G01N33/6854G01N2333/165G01N2469/20
Inventor 金腾川姜德华马欢常志远曾威红易如婷黄文喜
Owner GUANGZHOU KANGRUN BIOTECHNOLOGY CO LTD
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