Journal of Shandong University (Health Sciences) ›› 2023, Vol. 61 ›› Issue (4): 77-85.doi: 10.6040/j.issn.1671-7554.0.2022.0859

• 临床医学 • Previous Articles    

Preparation and application of an enzyme-linked immunoassay kit for SARS-CoV-2 neutralizing antibody

GAO Lihe1, REN Jingjing2, LI Yan3, LI Qiang4, MA Wanshan5, LI Huanjie6, CHEN Zhen7,OU Lanxiang1, ZHANG Shaoming1, ZHU Zhiwei1, DING Xinglong7, LI Hongxia7, WANG Yan7, ZHANG Zhongfa4, WANG Yunshan8   

  1. 1. Shandong Institute of Industrial Technology for Health Sciences and Precision Medicine, Jinan 250100, Shandong, China;
    2. Laboratory Pathology, 80th Army Hospital of PLA, Weifang 261021, Shandong, China;
    3. Institute for the Prevention and Control of Infectious Diseases, Shandong Center for Disease Control and Prevention, Jinan 250014, Shandong, China;
    4. Liver Disease Center, Shandong Public Health Clinical Center, Jinan 250102, Shandong, China;
    5. Department of Clinical Laboratory, The First Affiliated Hospital of Shandong First Medical University, Jinan 250013, Shandong, China;
    6. Medical Integration and Practice Center, Cheeloo College of Medicine, Shandong University, Jinan 250012, Shandong, China;
    7. Shandong Laibo Biotechnology Co. Ltd, Jinan 250101, Shandong, China;
    8. Medical Research &
    Laboratory Diagnostic Center, The Affiliated Central Hospital of Shandong First Medical University, Jinan 250013, Shandong, China
  • Published:2023-04-11

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Abstract: Objective To develop an enzyme-linked immunosorbent assay kit for the detection of SARS-CoV-2 neutralizing antibody, and to analyze its clinical value in monitoring the efficacy of COVID-19 vaccine. Methods After the receptor binding domain(RBD)of SARS-CoV-2 virus S protein and human angiotensin-converting enzyme 2(ACE2)were expressed and purified, RBD was conjugated with horseradish peroxidase(HRP)and ACE2 was conjugated with biotin(Bio)to prepare a SARS-CoV-2 neutralizing antibody detection kit. The serum samples of 15 volunteers injected with COVID-19 vaccine were collected to analyze the level of neutralizing antibody produced. Results The positive rate of SARS-CoV-2 neutralizing antibody was 0.00% before vaccination. On the 11th to 15th day after the first, second and third doses of COVID-19 vaccine injection, the positive rate of SARS-CoV-2 neutralizing antibody was 13.33%, 93.33% and 100.00%. On the 180th to 190th day after the second dose, the positive rate of neutralizing antibody reduced to 33.33%. The performance test showed the kit’s detection limit was 0.06 μg/mL, the coefficient of variation(CV)in the batch was <5%, CV between the batches was <10%, and the linear detection range was 0.030-3.125 μg/mL. Furthermore, the validation value for the analytical sensitivity was 0.040 μg/mL, which was highly consistent with the results of microcellular neutralization assay. Conclusion The enzyme-linked immunosorbent assay kit for SARS-CoV-2 neutralizing antibody has good performance, and the detection method is fast, convenient, and easy to operate, which can be used to evaluate the protective effects of the vaccines.

Key words: Enzyme-linked immunosorbent assay, SARS-CoV-2, Neutralizing antibody, COVID-19 vaccine, Microcellular neutralization assay

CLC Number: 

  • R563.1
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