A Comparative Study of Real-Time RT-PCR-Based SARS-CoV-2 Detection Methods and Its Application to Human-Derived and Surface Swabbed Material.

The Journal of Molecular Diagnostics : JMD
Aizhan TastanovaMitchell P Levesque

Abstract

Real-time RT-PCR remains a gold standard in the detection of various viral diseases. In the coronavirus 2019 pandemic, multiple RT-PCR-based tests were developed to screen for viral infection. As an emergency response to increasing testing demand, we established a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) PCR diagnostics platform for which we compared different commercial and in-house RT-PCR protocols. Four commercial, one customized, and one in-house RT-PCR protocols were evaluated with 92 SARS-CoV-2-positive and 92 SARS-CoV-2-negative samples. Furthermore, economical and practical characteristics of these protocols were compared. In addition, a highly sensitive digital droplet PCR (ddPCR) method was developed, and application of RT-PCR and ddPCR methods on SARS-CoV-2 environmental samples was examined. Very low limits of detection (1 or 2 viral copies/μL), high sensitivities (93.6% to 97.8%), and high specificities (98.7% to 100%) for the tested RT-PCR protocols were found. Furthermore, the feasibility of downscaling two of the commercial protocols, which could optimize testing capacity, was demonstrated. Tested commercial and customized RT-PCR detection kits show very good and comparable sensitivity and sp...Continue Reading

References

Jan 30, 2020·Euro Surveillance : Bulletin Européen Sur Les Maladies Transmissibles = European Communicable Disease Bulletin·Victor M CormanChristian Drosten
May 18, 2020·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Puck B van KasterenAdam Meijer
Jun 4, 2020·Annals of Internal Medicine·Daniel P Oran, Eric J Topol
Jun 23, 2020·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Zsófia IglóiRichard Molenkamp
Jul 7, 2020·The Lancet Infectious Diseases·Eskild PetersenLone Simonsen
Oct 9, 2020·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·Bo DiaoYuzhang Wu

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