Cautionary Note on Contamination of Reagents Used for Molecular Detection of SARS-CoV-2

Jim F Huggett; Vladimir Benes; Stephen A Bustin; Jeremy A Garson; Karthyn Harris; Martin Kammel; Mikael Kubista; Timothy D McHugh; Jacob Moran-Gilad; Tania Nolan; Michael W Pfaffl; Marc Salit; Greg Shipley; Peter M Vallone; Jo Vandesompele; Carl Wittwer; Heinz Zeichhardt

doi:10.1093/clinchem/hvaa214

Cautionary Note on Contamination of Reagents Used for Molecular Detection of SARS-CoV-2

Clin Chem. 2020 Nov 1;66(11):1369-1372. doi: 10.1093/clinchem/hvaa214.

Authors

Jim F Huggett^{1

2}, Vladimir Benes³, Stephen A Bustin⁴, Jeremy A Garson⁵, Karthyn Harris⁶, Martin Kammel⁷, Mikael Kubista⁸, Timothy D McHugh⁹, Jacob Moran-Gilad¹⁰, Tania Nolan¹¹, Michael W Pfaffl¹², Marc Salit^{13

14}, Greg Shipley¹⁵, Peter M Vallone¹⁶, Jo Vandesompele^{17

18}, Carl Wittwer¹⁹, Heinz Zeichhardt²⁰

Affiliations

¹ National Measurement Laboratory (NML) at LGC, Queens Rd, Teddington, UK.
² School of Biosciences & Medicine, Faculty of Health & Medical Science, University of Surrey, Guildford, UK.
³ Genomics Core Facility, EMBL Heidelberg, Heidelberg, Germany.
⁴ Medical Technology Research Centre, Faculty of Health, Social Care & Education, Anglia Ruskin University, Essex, UK.
⁵ Division of Infection and Immunity, University College London, London, UK.
⁶ Microbiology Department, Great Ormond Street Hospital NHS Foundation Trust, London, UK.
⁷ INSTAND e. V., Duesseldorf; IQVD GmbH, Institut fuer Qualitaetssicherung in der Virusdiagnostik, Berlin, Germany.
⁸ TATAA Biocenter, Sweden and Institute of Biotechnology of the Czech Academy of Sciences, Czech Republic.
⁹ Division of Infection and Immunity, Center for Clinical Microbiology, University College London, UK.
¹⁰ Department of Health Systems Management, School of Public Health, Faculty of Health Sciences, Ben Gurion University of the Negev, Beer Sheva, Israel.
¹¹ Institute of Population Health, Faculty of Medical and Human Sciences, University of Manchester, The Gene Team, Manchester, UK.
¹² Animal Physiology and Immunology, School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany.
¹³ Joint Initiative for Metrology in Biology, SLAC National Accelerator Laboratory, Menlo Park, CA.
¹⁴ Department of Bioengineering and Pathology, Stanford University, Stanford, CA.
¹⁵ Shipley Consulting, LLC, Vancouver, WA.
¹⁶ National Institute of Standards and Technology, Gaithersburg, MD.
¹⁷ Department of Biomolecular Medicine, Ghent University.
¹⁸ Biogazelle, Belgium; Ghent University, Belgium.
¹⁹ Department of Pathology, University of Utah, Salt Lake City, UT.
²⁰ INSTAND e. V., Duesseldorf; Charité - University Medicine Berlin; IQVD GmbH, Institut fuer Qualitaetssicherung in der Virusdiagnostik, Berlin, Germany.

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: RT-qPCR; SARS-CoV-2; contamination; false positive; molecular diagnosis.

MeSH terms

COVID-19 / diagnosis*
COVID-19 / virology
Diagnostic Errors
Humans
Indicators and Reagents / chemistry*
Limit of Detection
Nasopharynx / virology
RNA, Viral / genetics
RNA, Viral / metabolism
Reverse Transcriptase Polymerase Chain Reaction
SARS-CoV-2 / genetics*
SARS-CoV-2 / isolation & purification
Specimen Handling / standards

Substances

Indicators and Reagents
RNA, Viral