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Test strip for detecting novel coronavirus antibody, and preparation method and application of test strip

A coronavirus and test strip technology, applied in the field of colloidal gold test strips, can solve the problems of poor operation simplicity, cumbersome operation, and long time consumption

Active Publication Date: 2020-07-10
JIANGSU PROVINCIAL CENT FOR DISEASE CONTROL & PREVENTION PUBLIC HEALTH RES INST OF JIANGSU PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzyme-linked immunosorbent assay (ELISA), the most widely used serological detection method, has accurate results, but it is not easy to operate, requires multiple incubations and washings, is cumbersome to operate, and takes a long time. It takes at least 1.5 hours to detect the results

Method used

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  • Test strip for detecting novel coronavirus antibody, and preparation method and application of test strip
  • Test strip for detecting novel coronavirus antibody, and preparation method and application of test strip
  • Test strip for detecting novel coronavirus antibody, and preparation method and application of test strip

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Embodiment 1

[0079] Expression and purification of embodiment 1SARS-CoV-2NP protein

[0080] 1. Expression and purification of SARS-CoV-2NP protein

[0081] Take the inactivated SARS-CoV-2 strain, and use the QIAamp RNA mini kit (Qiagen, Germany) to prepare the viral RNA template. The open reading frame of SARS-CoV-2 protein was amplified using the Invitrogen One step RT-PCR kit (Invitrogen, USA); the reading frame was inserted into the bacterial expression plasmid pET-28(a)+ (Qiagen, Germany) using molecular biology methods , to obtain the bacterial expression plasmid pET-28(a)-NP. The plasmid was routinely transformed into E.CoLi BL21 strain (Qiagen, Germany), and protein was induced in LB broth at 37°C with 50 μg / ml kanamycin and 1 mM isopropyl-β-D-thiogalactopyranoside. Express. Cultured pellets containing recombinant proteins were resuspended in chromatography binding buffer and sonicated. The cell lysate was centrifuged, and the supernatant was used to load the nickel ion affinit...

Embodiment 2

[0093] The preparation of embodiment 2 colloidal gold immunochromatography test strips

[0094] 1. Preparation of test reagents

[0095] Gold-labeled antibody complex solution A: 0.01mol / L PBS solution (pH 7.4) containing 0.5% Tween-20 (v / v), 5% sucrose (w / v), and 1% BSA (w / v).

[0096] Gold-labeled antibody complex solution B: 0.8% Tris solution (pH 8.0) containing 0.5% Tween-20 (v / v), 2% sucrose (w / v), 0.1% BSA (w / v).

[0097] Conjugate pad treatment solution A: 0.01 mol / L PBS solution (pH 7.2) containing 0.1% Tween-20 (v / v), 5% sucrose (w / v), and 1% BSA (w / v).

[0098] Conjugate pad treatment solution B: 0.01 mol / L PBS solution (pH 7.2) containing 0.2% Tween-20 (v / v), 2% sucrose (w / v), and 1% BSA (w / v).

[0099] 2. Experimental method

[0100] 2.1 Preparation of colloidal gold particles

[0101] Add 1g of chloroauric acid into a clean glass bottle, then add ultrapure water to make up to 100mL, filter and sterilize with a 0.45μm filter; take 1mL of the prepared 1% chloro...

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Abstract

The invention discloses a test strip for detecting a novel coronavirus antibody, and a preparation method and application of the test strip. The colloidal gold immunochromatography test strip preparedby the method has good sensitivity, specificity and stability, is convenient to store, long in storage life, simple to operate and high in detection speed, is suitable for rapid diagnosis of SARS-CoV-2 virus infection, and can be used for in-situ detection of basic tissues.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a colloidal gold test strip for detecting novel coronavirus antibodies, a preparation method and application thereof. Background technique [0002] Severe acute respiratory syndrome coronavirus 2 (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2), a coronavirus belonging to the genus β, has an envelope, and the particles are round or oval, often pleomorphic, with a diameter of 60- 140nm. Patients infected with 2019-nCoV may be combined with multiple organ damages such as ARDS, myocardial damage, abnormal coagulation function, kidney damage, and liver damage. [0003] At present, nucleic acid detection is an important means of diagnosing 2019-nCoV infection. However, PCR technology requires huge equipment, time-consuming and labor-intensive detection, and high requirements for laboratory divisions and operators, making it difficult to meet the needs of emergency response to ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/569G01N33/558G01N33/532
CPCG01N33/532G01N33/558G01N33/56983G01N33/577G01N33/587G01N33/6854G01N2333/165G01N2469/20
Inventor 温恬黄超焦永军曾晓燕史凤娟朱宝立
Owner JIANGSU PROVINCIAL CENT FOR DISEASE CONTROL & PREVENTION PUBLIC HEALTH RES INST OF JIANGSU PROVINCE
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