A novel coronavirus antibody detection kit based on magnetic particle chemiluminescence

A coronavirus and antibody detection technology, applied in chemiluminescence/bioluminescence, analysis through chemical reaction of materials, measurement devices, etc., can solve the problems of prone to false negatives, low sensitivity, and prone to false positives

Active Publication Date: 2021-03-16
DYNAMIKER BIOTECH TIANJIN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, imaging diagnosis has the problem of different diseases with the same shadow, low specificity, and false positive results are prone to occur; molecular diagnosis has high specificity, but low sensitivity, and false negative results are prone to occur
For serological diagnostic methods, the colloidal gold method has high sensitivity, but low specificity, and is prone to false positives; enzyme-linked immunoassay requires manual operation, and there are biosafety risks

Method used

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  • A novel coronavirus antibody detection kit based on magnetic particle chemiluminescence
  • A novel coronavirus antibody detection kit based on magnetic particle chemiluminescence
  • A novel coronavirus antibody detection kit based on magnetic particle chemiluminescence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] This embodiment provides a novel coronavirus IgM antibody detection kit based on magnetic particle chemiluminescence, specifically including:

[0079] 0.3 mg / mL streptavidin magnetic particles, biotin-labeled novel coronavirus antigen, acridine sulfonamide-labeled goat anti-human IgM antibody, sample diluent, positive quality control, negative quality control, sample diluent and washing liquid.

[0080] Among them, (1) The preparation method of the biotin-labeled novel coronavirus antigen is:

[0081] Dilute 0.3 mg of the recombinant nucleocapsid protein of the new coronavirus and 0.2 mg of the recombinant spike protein S1 with 0.02 MPBS (pH7.2) buffer, and the final concentration of the mixed antigen is 1 mg / mL;

[0082] Wherein, the sequence of the recombinant nucleocapsid protein is SEQ ID NO.1; the sequence of the recombinant spike protein S1 is SEQ ID NO.3:

[0083] Take 12 μL of 10 mg / mL activated biotin and add it to the above antigen buffer, mix well, and keep...

Embodiment 2

[0090] This embodiment provides a new coronavirus IgM antibody detection kit based on magnetic particle chemiluminescence. The difference from Example 1 is that the mass ratio of recombinant nucleocapsid protein and recombinant spike protein S1 is set to 5:0, 4:0, respectively. 1, 2:3, 1:4 and 0:5;

[0091] Detection discrimination, the results are shown in Table 1:

[0092]

[0093] Among them, the reference samples are standard samples, which are the samples to be tested during the development of the kit, and are numbered N01~N15 (negative samples), P01~05 (positive samples) and L1~L3 (quality control samples); from the above table It can be seen that when the ratio of nucleocapsid protein and spike protein S1 is 5:0, 4:1, 1:4, and 0:5, the negative coincidence rate (that is, the discrimination degree is less than 1) is 14 / 15;

[0094] Specifically: when the ratio is 5:0, the discrimination of N10 samples is 2.08; when the ratio is 4:1, the discrimination of N10 samples ...

Embodiment 3

[0097] This example provides a new coronavirus IgM antibody detection kit based on magnetic particle chemiluminescence. The difference from Example 1 is that the addition amount of Sulfo-NHS-LC-Biotin solution is set to 3, 6 and 24 μL respectively.

[0098] The specific results are shown in Table 2 below;

[0099]

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Abstract

The invention provides a novel coronavirus antibody detection kit based on magnetic particle chemiluminescence. The detection kit includes: streptavidin magnetic particles, biotin-labeled novel coronavirus antigen, acridine sulfonamide-labeled secondary antibody, sample diluent and quality control; the biotin-labeled novel coronavirus Antigens include recombinant nucleocapsid protein and recombinant spike protein S1. Mix the sample to be tested, biotin-labeled antigen and streptavidin magnetic particles, incubate and wash, and then add acridine sulfonamide-labeled antibody to form magnetic particles-streptavidin-biotin-antigen-new coronavirus Virus antibody-secondary antibody complex, and then detect the luminescence intensity to achieve the qualitative of the sample to be tested.

Description

technical field [0001] The invention belongs to the technical field of immunoassay and detection, and relates to a novel coronavirus antibody detection kit based on magnetic particle chemiluminescence. Background technique [0002] Coronaviruses are a class of single-stranded positive-sense RNA viruses with an envelope (envelope), with a diameter of about 60-220 nm, and widely exist among humans and other mammals. Most coronavirus infections are mild, but two coronaviruses have had outbreaks with severe consequences: severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV). [0003] The new coronavirus (2019-nCoV) can be transmitted through respiratory droplets, contact, etc., and has a strong ability to infect humans. Its basic reproduction number R0 is about 2.2 (90% high-density interval 1.4-3.8). The most common symptoms of this viral infection were fever, cough, myalgia, or fatigue. All patients developed pneu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/532G01N21/76
CPCG01N33/56983G01N33/54326G01N33/532G01N21/76G01N2469/20G01N2333/165G01N33/54353G01N33/533G01N33/563
Inventor 刘春龙张舟马少华王家顺粟艳周泽奇
Owner DYNAMIKER BIOTECH TIANJIN
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