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Oxford University Press - PMC COVID-19 Collection logoLink to Oxford University Press - PMC COVID-19 Collection
. 2020 Sep 7:hvaa214. doi: 10.1093/clinchem/hvaa214

Cautionary note on contamination of reagents used for molecular detection of SARS-CoV-2

Jim F Huggett 1,2,, Vladimir Benes 3, Stephen A Bustin 4, Jeremy A Garson 5, Karthyn Harris 6, Martin Kammel 7, Mikael Kubista 8, Timothy D McHugh 9, Jacob Moran-Gilad 10, Tania Nolan 11, Michael W Pfaffl 12, Marc Salit 13,14, Greg Shipley 15, Peter M Vallone 16, Jo Vandesompele 17,18, Carl Wittwer 19, Heinz Zeichhardt 20
PMCID: PMC7499524  PMID: 32894756

Abstract

Reverse transcription (RT)-PCR, the principal diagnostic method applied in the world-wide struggle against COVID-19, is capable of detecting a single molecule of a viral genome. Correctly designed and practiced RT-PCR assays for SARS-CoV-2 should not cross react with similar but distinct viral pathogens, such as the coronaviruses associated with the common cold, and should perform with very high analytical sensitivity. This analytical performance is predicated on the ability of the method to detect the presence of the selected nucleic acid target, without detection of a false positive signal.

Keywords: SARS-CoV-2, RT-qPCR, Contamination, False positive, Molecular Diagnosis


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