Frontiers in Microbiology (Feb 2021)

An Optimized High-Throughput Immuno-Plaque Assay for SARS-CoV-2

  • Alberto A. Amarilla,
  • Naphak Modhiran,
  • Naphak Modhiran,
  • Yin Xiang Setoh,
  • Nias Y. G. Peng,
  • Julian D. J. Sng,
  • Benjamin Liang,
  • Christopher L. D. McMillan,
  • Morgan E. Freney,
  • Stacey T. M. Cheung,
  • Keith J. Chappell,
  • Keith J. Chappell,
  • Keith J. Chappell,
  • Alexander A. Khromykh,
  • Alexander A. Khromykh,
  • Paul R. Young,
  • Paul R. Young,
  • Paul R. Young,
  • Daniel Watterson,
  • Daniel Watterson,
  • Daniel Watterson

DOI
https://doi.org/10.3389/fmicb.2021.625136
Journal volume & issue
Vol. 12

Abstract

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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.

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