AUTHOR=Amarilla Alberto A. , Modhiran Naphak , Setoh Yin Xiang , Peng Nias Y. G. , Sng Julian D. J. , Liang Benjamin , McMillan Christopher L. D. , Freney Morgan E. , Cheung Stacey T. M. , Chappell Keith J. , Khromykh Alexander A. , Young Paul R. , Watterson Daniel TITLE=An Optimized High-Throughput Immuno-Plaque Assay for SARS-CoV-2 JOURNAL=Frontiers in Microbiology VOLUME=12 YEAR=2021 URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.625136 DOI=10.3389/fmicb.2021.625136 ISSN=1664-302X ABSTRACT=

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been identified as the causative agent of coronavirus disease 2019 and is capable of human-to-human transmission and rapid global spread. The rapid emergence and global spread of SARS-CoV-2 has encouraged the establishment of a rapid, sensitive, and reliable viral detection and quantification methodology. Here, we present an alternative assay, termed immuno-plaque assay (iPA), which utilizes a combination of plaque assay and immunofluorescence techniques. We have extensively optimized the conditions for SARS-CoV-2 infection and demonstrated the great flexibility of iPA detection using several antibodies and dual-probing with two distinct epitope-specific antibodies. In addition, we showed that iPA could be utilized for ultra-high-throughput viral titration and neutralization assay within 24 h and is amenable to a 384-well format. These advantages will significantly accelerate SARS-CoV-2 research outcomes during this pandemic period.