Development and Performance Evaluation of A Rapid IgM-IgG Combined Antibody Test for 2019 Novel Coronavirus Infection

doi:10.13523/j.cb.2006044

China Biotechnology

2020, Vol. 40

Issue (8): 1-9 DOI: 10.13523/j.cb.2006044

Development and Performance Evaluation of A Rapid IgM-IgG Combined Antibody Test for 2019 Novel Coronavirus Infection

ZHANG Sai¹,XIANG Le¹,LI Lin-hai²,LI Hui-jun³,WANG Gang¹,QIAN Chun-gen^4,^*(

)

1 Shenzhen YHLO Biotech Co., Ltd., Shenzhen 518116, China
2 General Hospital of Southern Theater Command, Guangzhou 510010, China
3 Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China
4 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 immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies against 2019 novel coronavirus (2019-nCoV) and to evaluate its clinical performance. Methods: The colloidal gold was prepared by trisodium citrate reduction. The receptor binding domain (RBD) of spike protein and nucleocapsid protein (NP) were used as marker antigen. The nitrocellulose membrane was coated with mouse anti human IgM monoclonal antibody and mouse anti human IgG monoclonal antibody, and the detection reagent was prepared by using dinitrophenol-bovine serum albumin (DNP-BSA) and rabbit anti DNP polyclonal antibody as independent quality control. By comparing the clinical coincidence rate of RBD protein and NP protein, the better antigen was selected to prepare the detection reagent, and the performance of cross reactivity, interference reactivity, accelerated stability, specificity and sensitivity of clinical diagnosis were evaluated. Results: The total coincidence rate of RBD protein was 98.48% (389/395), and that of NP protein was 89.11% (352/395). There were no cross reaction with antibody positive samples of 13 common pathogens. Triglyceride, hemoglobin, bilirubin, rheumatoid factor (RF), human anti mouse antibody (HAMA) and antinuclear antibody (ANA) in the samples did not interfere with the test results. The kit was stable after 6 weeks accelerated at 50℃. The sensitivity of IgM was 78.31% (65/83), the specificity was 98.90% (721/729), the sensitivity of IgG was 92.77% (77/83), the specificity was 99.31% (724/729), the sensitivity of IgM and IgG combined detection was 92.77% (77/83), the specificity was 98.35% (717/729), the kappa consistency test had a kappa value of 0.883 0 (P<0.05). Conclusion: The 2019-nCoV IgM/IgG antibody detection reagent (colloidal gold method) has the advantages of high specificity and sensitivity, fast detection speed and portable operation, which can be used as a supplementary method for the existing 2019-nCoV nucleic acid detection method.

Key words： Novel coronavirus Antibodies Colloidal gold Immunochromatography

Received: 23 June 2020 Published: 10 September 2020

ZTFLH:

Q819

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

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	Sai ZHANG
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	Lin-hai LI
	Hui-jun LI
	Gang WANG
	Chun-gen QIAN

Cite this article:

ZHANG Sai,XIANG Le,LI Lin-hai,LI Hui-jun,WANG Gang,QIAN Chun-gen. Development and Performance Evaluation of A Rapid IgM-IgG Combined Antibody Test for 2019 Novel Coronavirus Infection. China Biotechnology, 2020, 40(8): 1-9.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2006044 OR https://manu60.magtech.com.cn/biotech/Y2020/V40/I8/1

Fig.1 Structure chart of test strip for IgM and IgG antibody combined 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

Table 2 Comparison of sensitivity and specificity between NP protein and RBD protein

Table 3 The results of accelerated stability

Table 4 The results of cross reaction

Table 5 The results of interference response

Table 6 Statistical results of clinical performance

Fig.3 The legend of clinical test The legend of clinical test (the upper picture shows the positive sample, and the lower picture shows the negative sample)


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