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Early screening method for new coronavirus

A coronavirus, a new type of technology, applied in the field of immune detection, can solve the problems of increased detection value, long window period, and inability to meet the needs of patients screened out, and achieve accurate detection results

Active Publication Date: 2021-07-13
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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  • Early screening method for new coronavirus
  • Early screening method for new coronavirus
  • Early screening method for new coronavirus

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): Tris buffer containing magnetic bead coating coated with SARS-CoV-2 recombinant antigen, pH value is 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); Antibody of interest in the assay.

[0039] 2. Anti-human IgA antibody acridinium ester marker (R2): 2-(N-morpholine) ethanesulfonic acid buffer solution for acridinium 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 emit light after being excited.

[0040] 3. Reaction buffer (R3): phosphate buffer co...

Embodiment 2

[0048] A test kit for screening novel coronavirus infection for detection of SARS-CoV-2 IgA antibody, 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 The Spike protein (S protein) expressed in the cultured human embryonic kidney cell line HEK293F, referred to as the S protein, truncated the RBD domain Gln321-Ser591 amino acid sequence for protein expression. The protein expression process is: connect the SARS-CoV-2 S protein RBD gene to human IgG1 Fc through a TEV restriction site sequence at the C-terminus, then clone it into the mammalian expression vector pTT5 and add the Kozak sequence in the translation initiation region , and then transfected into suspension culture cells HEK293F, using interferon α-1 as a signal peptide to make HEK293F cells secrete RBD-TEV-Fc fusion protein, and then using Protein A column and nickel column for two purifications; the first purification wa...

Embodiment 3

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

[0066] The kit for screening novel coronavirus infection includes:

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

[0068] 2. Anti-human IgA antibody acridinium ester marker (R2): 2-(N-morpholine) ethanesulfonic acid buffer solution for acridinium 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 c...

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Abstract

An earlier screening method for novel coronaviruses of the present invention can detect novel coronaviruses, especially for early screening of SARS‑CoV‑2 IgA antibodies. The kit contains, as R1, the SARS-CoV-2 magnetic bead coating: the magnetic bead coating tris buffer containing the SARS-CoV-2 recombinant antigen, the pH value is 7.1-7.4; Anti-human IgA antibody acridinium ester marker as R2: 2-(N-morpholine)ethanesulfonic acid buffer containing acridinium ester-labeled mouse anti-human IgA monoclonal antibody. The kit and SARS-CoV-2 IgA antibody immunoassay reagent of the present invention can be used for supplementary diagnosis of new coronary pneumonia at an earlier stage, and has the characteristics of accurate detection results.

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 Patents(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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