Method for preparing rapid detection test paper of novel coronavirus IgA antibody

A coronavirus and test strip technology, applied in the field of recombinant protein detection of novel coronavirus 2019-nCoV IgA antibodies, can solve the problems of lack of non-invasive, rapid and accurate screening of 2019 novel coronavirus infection, patient discomfort, cross-infection, etc. Achieve the effect of high-throughput detection, low detection environment requirements, and high fault tolerance.

Inactive Publication Date: 2020-06-05
BEIJING DIAGREAT BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, blood samples are invasive samples, which require professional medical staff to collect blood, and cause discomfort to patients, and the sampling process may cause unnecessary cross-infection
Therefore, there is a lack of non-invasive, rapid and accurate detection reagents for 2019 novel coronavirus infection on the market

Method used

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  • Method for preparing rapid detection test paper of novel coronavirus IgA antibody

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preparation example Construction

[0037] The invention provides the preparation method of described test strip, comprises the following steps:

[0038] 1) After the fluorescent microspheres are activated, they are mixed and coupled with the recombinant protein to obtain the 2019 novel coronavirus-specific antigen labeled with fluorescent microspheres;

[0039] 2) spraying the 2019 novel coronavirus-specific antigen labeled with fluorescent microspheres onto the glass fiber membrane to obtain a fluorescent microsphere pad;

[0040] 3) Anti-human IgA antibody and goat anti-chicken IgY antibody are sprayed respectively on the detection zone and the quality control zone of the nitrocellulose membrane to obtain the nitrocellulose membrane fixed with the detection zone and the quality control zone;

[0041] (4) Paste sample absorbent pads, fluorescent microsphere pads, nitrocellulose membranes fixed with detection areas and quality control areas, and water absorbent pads on the base plate in order to obtain test str...

Embodiment 1

[0052] Preparation of the novel coronavirus 2019-nCoV IgA antibody detection reagent strip.

[0053] 1. Preparation of novel coronavirus 2019-nCoV recombinant NP protein:

[0054] The new coronavirus 2019-nCoV recombinant NP protein has a cysteine ​​residue added to the C-terminus, and the amino acid sequence is shown in the sequence Seq ID No.1. Entrusted Beijing Deoping Biotechnology Co., Ltd. to produce and prepare, the purity is greater than 95%.

[0055]MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGLP NNTASWFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRG GDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKD HIGTRNPANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSR NSTPGSSRGTSPARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQG QTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDD KDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQ TVTLLPAADLDDFSKQLQQSMSSADSTQAGSSC

[0056] 2. Preparation of 2019-nCoV recombinant NP antigen labeled with fluorescent microsphere...

Embodiment 2

[0068] Application of time-resolved fluorescent immunochromatographic test strips for detection of novel coronavirus 2019-nCoV IgA antibody

[0069] 1. Sample pretreatment

[0070] Aspirate 20 μL saliva sample and add to 200 μL sample diluent, mix well.

[0071] 2. Test with test strips

[0072] Use a micropipette to accurately draw 80 μL of the sample solution to be tested into the sample hole of the test strip, and act for 15 minutes at room temperature (20-25°C); insert the test paper card into the carrier of the fluorescence detector, and select the sample to be tested through the touch screen. item, press the "Start Detection" button, the fluorescence detector will automatically scan the test paper card; read or print the test results on the display screen of the instrument.

[0073] 3. Analysis of test results

[0074] Quantitative detection

[0075] After the test is completed, the instrument obtains the ratio of the time-resolved fluorescence intensity of the detec...

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Abstract

The invention provides a recombinant protein and test paper for detecting a novel coronavirus 2019-nCoV IgA antibody, and a preparation method and application, and belongs to the technical field of virus detection. The amino acid sequence of the recombinant protein is shown in SEQ ID No.1; the test paper comprises a bottom plate as well as a sample absorption pad, a fluorescent microsphere pad, anitrocellulose membrane and a water absorption pad which are in sequential lap joint and adhesion to the bottom plate; a 2019 novel coronavirus specific antigen marked by fluorescent microspheres is sprayed to the fluorescent microsphere pad; a detection zone and a quality control zone are fixed on the nitrocellulose membrane; an anti-human IgA antibody is sprayed to the detection zone; and a sheep anti-chicken IgY antibody is sprayed to the quality control zone. By detecting the IgA antibody in a saliva sample, the test paper provided by the invention is capable of simply, rapidly and accurate detecting novel coronaviruses, and early-stage detection on the novel coronaviruses can be achieved.

Description

technical field [0001] The invention belongs to the technical field of virus detection, and in particular relates to a recombinant protein for detecting 2019-nCoV IgA antibody, a test strip, a preparation method and an application. Background technique [0002] Coronaviruses are a large family of viruses known to cause more serious illnesses such as colds, Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). Novel coronavirus is a new strain of coronavirus that has never been found in humans before, and 2019-nCoV is the seventh type of coronavirus isolated from humans. According to relevant disease monitoring, many cases of viral pneumonia were found, all of which were diagnosed as viral pneumonia / lung infection. [0003] Existing evidence shows that the 2019 novel coronavirus is mainly transmitted through droplets, aerosols, and direct contact with secretions, and can be transmitted from person to person with strong transmission power. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165G01N33/569G01N33/558G01N33/543G01N33/546
CPCC07K14/005C12N2770/20022G01N33/54313G01N33/558G01N33/56983
Inventor 周裕军周建平何昆仑田亚平
Owner BEIJING DIAGREAT BIOTECH CO LTD
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