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

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.

SUBMITTER: Amarilla AA

PROVIDER: S-EPMC7906992 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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

Amarilla Alberto A AA Modhiran Naphak N Setoh Yin Xiang YX Peng Nias Y G NYG Sng Julian D J JDJ Liang Benjamin B McMillan Christopher L D CLD Freney Morgan E ME Cheung Stacey T M STM Chappell Keith J KJ Khromykh Alexander A AA Young Paul R PR Watterson Daniel D

Frontiers in microbiology 20210212

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 immuno ...[more]

PMID: 33643253

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Dataset Information

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

Publications

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

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