Development of an automated, high-throughput SARS-CoV-2 neutralization assay based on a pseudotyped virus using a vesicular stomatitis virus (VSV) vector

作者：Ziteng Liang, Xi Wu, Jiajing Wu, Shuo Liu, Jincheng Tong, Tao Li, Yuanling Yu, Li Zhang, Chenyan Zhao, Qiong Lu, Haiyang Qin, Jianhui Nie, Weijin Huang, Youchun Wang · 发表于：Emerging Microbes & Infections · 年份：2023 · DOI：10.1080/22221751.2023.2261566 · 被引用次数：8 · 研究领域：SARS-CoV-2 and COVID-19 Research、SARS-CoV-2 detection and testing、Respiratory viral infections research

The global outbreak of COVID-19 has caused a severe threat to human health; therefore, simple, high-throughput neutralization assays are desirable for developing vaccines and drugs against COVID-19. In this study, a high-titre SARS-CoV-2 pseudovirus was successfully packaged by truncating the C-terminus of the SARS-CoV-2 spike protein by 21 amino acids and infecting 293 T cells that had been stably transfected with the angiotensin-converting enzyme 2 (ACE2) receptor and furin (named AF cells), to establish a simple, high-throughput, and automated 384-well plate neutralization assay. The method was optimized for cell amount, virus inoculation, incubation time, and detection time. The automated assay showed good sensitivity, accuracy, reproducibility, Z' factor, and a good correlation with the live virus neutralization assay. The high-throughput approach would make it available for the SARS-CoV-2 neutralization test in large-scale clinical trials and seroepidemiological surveys which would aid the accelerated vaccine development and evaluation.