Updated and validated pan-coronavirus pcr assay to detect all coronavirus genera

Myndi G. Holbrook, Simon J. Anthony, Isamara Navarrete-Macias, Theo Bestebroer, Vincent J. Munster, Neeltje van Doremalen*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

8 Citations (Scopus)
11 Downloads (Pure)


Coronavirus (CoV) spillover events from wildlife reservoirs can result in mild to severe human respiratory illness. These spillover events underlie the importance of detecting known and novel CoVs circulating in reservoir host species and determining CoV prevalence and distribution, allowing improved prediction of spillover events or where a human–reservoir interface should be closely monitored. To increase the likelihood of detecting all circulating genera and strains, we have modified primers published by Watanabe et al. in 2010 to generate a semi-nested pan-CoV PCR assay. Representatives from the four coronavirus genera (α-CoVs, β-CoVs, γ-CoVs and δ-CoVs) were tested and all of the in-house CoVs were detected using this assay. After comparing both assays, we found that the updated assay reliably detected viruses in all genera of CoVs with high sensitivity, whereas the sensitivity of the original assay was lower. Our updated PCR assay is an important tool to detect, monitor and track CoVs to enhance viral surveillance in reservoir hosts.

Original languageEnglish
Article number599
Issue number4
Publication statusPublished - 1 Apr 2021

Bibliographical note

Funding Information:
This research was funded by the Intramural Research Program, NIAID, NIH (1ZIAAI001179-01).

Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.


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