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Colloidal gold test strip for detecting 2019-nCoV antibody and preparation method thereof

A 2019-ncov, colloidal gold test paper technology, applied in the field of virus microorganisms, to achieve the effect of simple operation

Pending Publication Date: 2021-05-07
GENERAL HOSPITAL OF NINGXIA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] This application provides a colloidal gold test strip and preparation method for detecting 2019-nCoV antibodies, to solve the limitations of 2019-nCoV virus culture and isolation that require technicians to process samples for a long time and highly specialized skills question

Method used

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  • Colloidal gold test strip for detecting 2019-nCoV antibody and preparation method thereof
  • Colloidal gold test strip for detecting 2019-nCoV antibody and preparation method thereof

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preparation example Construction

[0038] On the one hand, the application provides a method for preparing a colloidal gold test strip for detecting 2019-nCoV antibodies, comprising the following steps:

[0039] After the chloroauric acid solution is heated, trisodium citrate is added to prepare a colloidal gold solution;

[0040] Add the colloidal gold solution to K 2 CO 3, anti-2019-nCoV monoclonal antibody and polyethylene glycol to prepare 2019-nCoV immune colloidal gold solution;

[0041] After the 2019-nCoV immune colloidal gold solution is pretreated, the solution is sprayed on the polyester film marking pad with a gold spraying machine at a gold spraying amount of 2.3 μL / cm to form a 2019-nCoV immune colloidal gold pad;

[0042] After the anti-2019-nCoV monoclonal antibody is pretreated, use a scribe machine to spot the nitrocellulose membrane detection line;

[0043] After pretreatment with goat anti-mouse IgG, spot the quality control line on the nitrocellulose membrane with a film scribe to obtain...

Embodiment

[0064] 1. Preparation of colloidal gold immune test strips for detection of 2019-nCoV

[0065] (1) Preparation of colloidal gold solution: 50mL mass fraction of 0.01% chloroauric acid is heated and boiled, and 0.55mL mass fraction of 1% trisodium citrate is added and mixed well, and then kept boiling for 30min, chloroauric acid is reduced into colloidal gold particles solution;

[0066] (2) Preparation of 2019-nCoV immune colloidal gold solution: take 50mL of the colloidal gold solution, and use 0.1M K 2 CO 3 Adjust its pH value to 7.8, add 2 mL of 1 mg / mL anti-2019-nCoV monoclonal antibody while stirring, and stir for 30 minutes; then add 1 mL of 25M polyethylene glycol with a molecular weight of 20,000 (PEG solution) dropwise, and stir for another 15 minutes; Centrifuge at 4°C and 20,000rpm for 15min, discard the supernatant, add 10mL of PBS buffer containing 0.4M PEG 20000 and pH=7.4 to suspend, centrifuge at 4°C and 20,000rpm for 15min, remove the supernatant, and repeat...

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Abstract

The invention provides a colloidal gold test strip for detecting a 2019-nCoV antibody and a preparation method thereof. The method comprises the following steps: heating a chloroauric acid solution, and adding trisodium citrate to prepare a colloidal gold solution; adding K2CO3, an anti-2019-nCoV monoclonal antibody and polyethylene glycol into the colloidal gold solution, and preparing a 2019-nCoV immune colloidal gold solution; pretreating the 2019-nCoV immune colloidal gold solution, and spraying the solution on a polyester film marking pad by using a metal spraying machine according to a metal spraying amount of 2.3 [mu] L / cm to form a 2019-nCoV immune colloidal gold pad; pretreating an anti-2019-nCoV monoclonal antibody, and applying the monoclonal antibody on a nitrocellulose membrane detection line by using a membrane scratching machine; pretreating goat anti-mouse IgG, and applying the pretreated goat anti-mouse IgG on a nitrocellulose membrane quality control line by using a membrane scratching machine to obtain a nitrocellulose membrane; and sequentially adhering the sample pad made of water-absorbing filter paper, the 2019-nCoV immune colloidal gold pad, the nitrocellulose membrane and the water-absorbing filter paper to a PVC rubber plate to be assembled into the test strip.

Description

technical field [0001] The present application relates to the technical field of viruses and microorganisms, in particular to a colloidal gold test strip for detecting 2019-nCoV antibodies and a preparation method. Background technique [0002] A novel coronavirus was subsequently identified as the causative agent, tentatively named 2019 novel coronavirus (2019-nCoV). With the emergence of 2019-nCoV, real-time RT-PCR is still the main means of diagnosing new virus strains in many available diagnostic platforms, and the time and type of RT-PCR specimen collection are of great significance to the diagnosis of 2019-nCoV. It was found that the respiratory specimens were positive for the virus in the early stage, but the serum was negative. It also showed that early in the disease, patients had high levels of the virus despite mild symptoms. In addition to the currently commonly used RT-PCR method for the diagnosis of MERS-CoV, four studies identified different diagnostic metho...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/68G01N33/558G01N33/543G01N33/532
CPCG01N33/532G01N33/543G01N33/558G01N33/56983G01N33/6854G01N2333/165G01N2469/20
Inventor 赵志军周云花杨宁爱康宇婷
Owner GENERAL HOSPITAL OF NINGXIA MEDICAL UNIV