Development and Performance Evaluation of a Rapid Antigen Test for SARS-CoV-2

doi:10.13523/j.cb.2102025

China Biotechnology

2021, Vol. 41

Issue (5): 27-34 DOI: 10.13523/j.cb.2102025

Development and Performance Evaluation of a Rapid Antigen Test for SARS-CoV-2

ZHANG Sai¹,WANG Gang¹,LIU Zhong-ming²,LI Hui-jun³,WANG Da-ming⁴,QIAN Chun-gen^5,^**(

)

1 Shenzhen YHLO Biotech Co., Ltd., Shenzhen 518116, China
2 General Hospital of Southern Theater Command, Guangzhou 510010, China
3 Tongji Hospital affiliated to Huazhong University of Science and Technology, Wuhan 430030, China
4 Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China
5 College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China

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Abstract

Objective: To establish a colloidal gold technique assay for the rapid detection of antigen against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and to evaluate its clinical performance. Methods: The colloidal gold was prepared by trisodium citrate reduction. The monoclonal antibody of mouse anti-SARS-CoV-2 nucleocapsid protein (NP) and dinitrophenol-bovine serum albumin (DNP-BSA) were labeled with colloidal gold nanoparticles. The monoclonal antibody of mouse anti-nucleocapsid protein and rabbit anti DNP polyclonal antibody were coated on the nitrocellulose membrane as detection line and quality control line to prepare immunofluorescence test strip. The performance of the limit of detection, cross-reactivity, accelerated stability, sensitivity and specificity of clinical diagnosis were evaluated. Results: The limit of detection for heat inactivated SARS-CoV-2 was 2.0×10² TCID₅₀/mL. There were no cross reaction with high concentration samples or cultured virus of 16 common pathogens. The kit was stable after 8 weeks accelerated at 50℃. Nasopharyngeal swab samples of clinical and healthy people were tested, the sensitivity was 96.67% (29/30), the specificity was 99.23% (129/130), the total coincidence rate was 98.75% (158/160), and the Kappa consistency test had a Kappa value of 0.959 0 (P<0.05). Conclusion: The SARS-CoV-2 antigen detection reagent (colloidal gold method) has the advantages of high sensitivity and specificity, fast detection speed, portable operation, no need for equipment and naked eye observation, which can be used as a supplementary method for the existing SARS-CoV-2 nucleic acid detection method.

Key words： SARS-CoV-2 Antigen Colloidal gold Immunochromatography Performance evaluation

Received: 22 February 2021 Published: 01 June 2021

ZTFLH:

Q816

Corresponding Authors: Chun-gen QIAN E-mail: chungen_qian@hust.edu.cn

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	Sai ZHANG
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Cite this article:

ZHANG Sai,WANG Gang,LIU Zhong-ming,LI Hui-jun,WANG Da-ming,QIAN Chun-gen. Development and Performance Evaluation of a Rapid Antigen Test for SARS-CoV-2. China Biotechnology, 2021, 41(5): 27-34.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2102025 OR https://manu60.magtech.com.cn/biotech/Y2021/V41/I5/27

Fig.1 Structure chart of test strip for antigen detection

Table 1 Particle size and zeta potential of colloidal gold and its coupling

Fig.2 Particle size distribution of colloidal gold and its coupling (a)Unlabeled colloidal gold (b) Mouse anti-N protein antibody conjugate (c) DNP-BSA protein conjugate

Fig.3 The color intensities of different concentrations of inactivated culture.

Table 2 The results of accelerated stability

Table 3 The results of cross reaction

Table 4 The results of clinical performance evaluation


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