期刊
FRONTIERS IN MICROBIOLOGY
卷 12, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fmicb.2021.625136
关键词
coronaviruses; SARS-CoV-2; immuno-plaque assay (iPA); viral quantification
类别
资金
- Medical Research Future Fund (MRFF Novel Coronavirus Vaccine Development Grant) [APP1202445-2020]
The article introduces the iPA detection method, utilizing multiple antibodies and dual-specificity probe antibodies for detecting SARS-CoV-2, and demonstrates its use for high-throughput viral titration and neutralization assays within 24 hours, with compatibility in a 384-well format.
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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