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Novel coronavirus antibody magnetic immunochemiluminescence detection kit and application thereof

A chemiluminescence detection, coronavirus technology, applied in the field of immunological detection, can solve the problems of low detection limit and accuracy, low specificity of new coronavirus antibody detection, etc., to reduce human operation errors, shorten detection time, The effect of improving the detection throughput

Pending Publication Date: 2022-03-01
HUAZHONG UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention solves the technical problems of low specificity, low detection limit and low accuracy in the detection of novel coronavirus antibodies in the prior art

Method used

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  • Novel coronavirus antibody magnetic immunochemiluminescence detection kit and application thereof
  • Novel coronavirus antibody magnetic immunochemiluminescence detection kit and application thereof
  • Novel coronavirus antibody magnetic immunochemiluminescence detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: Preparation of specific components of the kit

[0028] 1. Preparation of enzyme-labeled antibody

[0029] A. Alkaline phosphatase activation

[0030] Use DMSO as solvent to prepare 10mM SMCC solution; add 240uL 5mg / mL alkaline phosphatase to 29.5uL 10mM SMCC solution, mix well and let stand at room temperature for 15min, then add 2.7uL 1M pH 7.3 glycine buffer solution, stand at room temperature for 5min ; 0.01M PBS solution with pH 7.4 is desalted by ultrafiltration, and the absorbance at 280nm is measured. The enzyme concentration is: ABS280*dilution factor / 1.0, the enzyme quality: enzyme concentration*product volume (mg / mL), and stored at 2-8°C.

[0031] B. Goat anti-human secondary antibody activation

[0032] Prepare 5mM EDTA.2H at pH 8.0 with 0.1M borate as solvent 2 O solution; take 400uL of 3mg / mL monoclonal antibody, use the above solution as the mobile phase, pass through the PD10 desalting column of the protein separation and purification sys...

Embodiment 2

[0035] Embodiment 2: Novel coronavirus IgM detection kit

[0036] A magnetic immunochemiluminescence detection kit for detecting novel coronavirus IgM antibodies was set up to contain the following components:

[0037] (1) Goat anti-human IgM secondary antibody labeled with alkaline phosphatase

[0038] (2) Magnetically labeled fluorescein isothiocyanate (FITC) antibody

[0039] (3) Antigen labeled with fluorescein isothiocyanate (FITC)

[0040] (4) Novel coronavirus IgM antibody calibration solution

[0041] (5) Positive control solution

[0042] (6) Negative control solution

[0043] (7) Sample diluent

[0044] (8) Chemiluminescence substrate solution

[0045] (9) Concentrated washing solution

[0046] The enzyme-labeled antibody diluent used in the present invention is 0.5M Tris buffer containing 5% bovine serum albumin (BSA), pH 7.5.

[0047] The magnetic label fluorescein isothiocyanate (FITC) antibody diluent is pH 8.0, containing 0.02% WPS (V:V), 1% TritionX-100 (...

Embodiment 3

[0055] Embodiment 3: Novel coronavirus IgG detection kit

[0056] A magnetic immunochemiluminescent detection kit for detecting novel coronavirus IgG antibodies was set up so that it contained the following components:

[0057] (1) Goat anti-human IgG secondary antibody labeled with alkaline phosphatase

[0058] (2) Magnetically labeled fluorescein isothiocyanate (FITC) antibody

[0059] (3) Antigen labeled with fluorescein isothiocyanate (FITC)

[0060] (4) Novel coronavirus IgG antibody calibration solution

[0061] (5) Positive control solution

[0062] (6) Negative control solution

[0063] (7) Sample diluent

[0064] (8) Chemiluminescence substrate solution

[0065] (9) Concentrated washing solution

[0066] The enzyme-labeled antibody diluent used in the present invention is 0.5M Tris buffer containing 5% bovine serum albumin (BSA), pH 7.5.

[0067] The magnetic label fluorescein isothiocyanate (FITC) antibody diluent is pH 8.0, containing 0.02% WPS (V:V), 1% Tri...

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Abstract

The invention relates to a novel coronavirus antibody magnetic immunochemiluminescence detection kit and application, and belongs to the field of immunological detection. The kit comprises the following reagents: an enzyme labeled secondary antibody, a magnetic labeled fluorescein isothiocyanate antibody, a fluorescein isothiocyanate modified novel coronavirus antigen, a calibration solution, a positive reference substance, a negative reference substance and a sample diluent, the novel coronavirus antigen on the fluorescein isothiocyanate modified novel coronavirus antigen can be specifically combined with a novel coronavirus antibody; the magnetic labeled fluorescein isothiocyanate antibody can be specifically combined with the fluorescein isothiocyanate on the novel coronavirus antigen modified by the fluorescein isothiocyanate; the enzyme-labeled secondary antibody can be specifically combined with a new coronavirus antibody; enzyme on the enzyme-labeled secondary antibody can degrade a luminous substrate in the luminous substrate liquid, so that the luminous substrate emits light. When the kit is used for detecting the antibody, the effects of relatively high detection limit, specificity and accuracy are achieved.

Description

technical field [0001] The invention belongs to the field of immunological detection, and in particular relates to a novel coronavirus antibody magnetic immunochemiluminescent detection kit and its application. Background technique [0002] The International Committee on Taxonomy of Viruses (ICTV) announced on February 11, 2020 that the official classification of the new coronavirus (2019-nCoV) was named severe acute respiratory syndrome coronavirus 2 (severe acute respiratory syndrome coronavirus 2). 2, SARS-CoV-2). The World Health Organization (WHO) announced on the same day that the official name of the disease caused by the virus is COVID-19. Coronaviruses are a large family of viruses known to cause colds as well as more serious illnesses such as Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). The novel coronavirus is a new strain of coronavirus that has never been found in humans before. [0003] After a person is infected with...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/535G01N33/533
CPCG01N33/56983G01N33/533G01N33/535G01N33/54326G01N2469/20G01N2333/165
Inventor 杨祥良杨海张德军杨宝凤金伟宋谦
Owner HUAZHONG UNIV OF SCI & TECH
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