Establishment of replication-competent vesicular stomatitis virus-based recombinant viruses suitable for SARS-CoV-2 entry and neutralization assays

Abstract

Replication-competent vesicular stomatitis virus (VSV)-based recombinant viruses are useful tools for studying emerging and highly pathogenic enveloped viruses in level 2 biosafety facilities. Here, we used a replication-competent recombinant VSVs (rVSVs) encoding the spike (S) protein of SARS-CoV-2 in place of the original G glycoprotein (rVSV-eGFP-SARS-CoV-2) to develop a high-throughput entry assay for SARS-CoV-2. The S protein was incorporated into the recovered rVSV-eGFP-SARS-CoV-2 particles, which could be neutralized by sera from convalescent COVID-19 patients. The recombinant SARS-CoV-2 also displayed entry characteristics similar to the wild type virus, such as cell tropism and pH-dependence. The neutralizing titers of antibodies and sera measured by rVSV-eGFP-SARS-CoV-2 were highly correlated with those measured by wild-type viruses or pseudoviruses. Therefore, this is a safe and convenient screening tool for SARS-CoV-2, and it may promote the development of COVID-19 vaccines and therapeutics.

Neutralizing activity of Sputnik V vaccine sera against SARS-CoV-2 variants

“…Several groups have now generated replication-competent VSV expressing SARS-CoV-2 spike in place of VSV-G (rcVSV-CoV2-S)( Dieterle et al, 2020 ; Li et al, 2020 )( Case, Rothlauf, Chen, Liu, et al, 2020 )( Baum et al, 2020 ; Weisblum et al, 2020 )( Yahalom-Ronen et al, 2020 )( Baum et al, 2020 ). These rcVSV-CoV2-S can be used in BSL-2 compatible virus neutralization assays (VNAs), which correlate very well with VNAs using live SARS-CoV-2 (Spearman’s r > 0.9 across multiple studies).…”

Construction and applications of SARS-CoV-2 pseudoviruses: a mini review

“…Along with the aforementioned study in Letko et al, Hoffmann and colleagues used pseudotyped SARS-CoV-2 particles to show that SARS-CoV-2 infection depends on the host cell factors ACE2 and transmembrane protease, serine 2 (TMPRSS2) [36]. Li et al recovered rVSV-eGFP-SARS-CoV-2 particles and confirmed that the pseudovirus displayed entry characteristics similar to that of the WT virus, such as cell tropism and pH-dependence [48]. Ou et al used a lentiviral pseudotype system to determine cell type susceptibility, virus receptor, and entry pathway for SARS-CoV-2 [23].…”

Development of an ultrasensitive fluorescent immunochromatographic assay based on multilayer quantum dot nanobead for simultaneous detection of SARS-CoV-2 antigen and influenza A virus

“…Influenza A (H1N1 2009/A) was supplied by our laboratory, which was cultured in chick embryo at 35 °C for 2 days, then quantified by plaque assay, and frozen at -70 °C until use [ 36 ]. Inactivated SARS-CoV-2 virions (∼10 8 pfu/mL) were provided by courtesy of Prof. Chengfeng Qin’s laboratory in the Beijing Institute of Microbiology and Epidemiology (Beijing, China) [ [37] , [38] , [39] ]. Human throat swab specimens were obtained from healthy volunteers from Beijing Institute of Radiation Medicine (Beijing, China), with the approval of Ethics Committee of the Institute.…”