Analytical sensitivity and efficiency comparisons of SARS-CoV-2 RT–qPCR primer–probe sets

Chantal B.F. Vogels, Anderson F. Brito, Anne L. Wyllie, Joseph R. Fauver, Isabel M. Ott, Chaney C. Kalinich, Mary E. Petrone, Arnau Casanovas-Massana, M. Catherine Muenker, Adam J. Moore, Jonathan Klein, Peiwen Lu, Alice Lu-Culligan, Xiaodong Jiang, Daniel J. Kim, Eriko Kudo, Tianyang Mao, Miyu Moriyama, Ji Eun Oh, Annsea ParkJulio Silva, Eric Song, Takehiro Takahashi, Manabu Taura, Maria Tokuyama, Arvind Venkataraman, Orr El Weizman, Patrick Wong, Yexin Yang, Nagarjuna R. Cheemarla, Elizabeth B. White, Sarah Lapidus, Rebecca Earnest, Bertie Geng, Pavithra Vijayakumar, Camila Odio, John Fournier, Santos Bermejo, Shelli Farhadian, Charles S. Dela Cruz, Akiko Iwasaki, Albert I. Ko, Marie L. Landry, Ellen F. Foxman, Nathan D. Grubaugh

Research output: Contribution to journalArticlepeer-review

525 Scopus citations

Abstract

The recent spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) exemplifies the critical need for accurate and rapid diagnostic assays to prompt clinical and public health interventions. Currently, several quantitative reverse transcription–PCR (RT–qPCR) assays are being used by clinical, research and public health laboratories. However, it is currently unclear whether results from different tests are comparable. Our goal was to make independent evaluations of primer–probe sets used in four common SARS-CoV-2 diagnostic assays. From our comparisons of RT–qPCR analytical efficiency and sensitivity, we show that all primer–probe sets can be used to detect SARS-CoV-2 at 500 viral RNA copies per reaction. The exception for this is the RdRp-SARSr (Charité) confirmatory primer–probe set which has low sensitivity, probably due to a mismatch to circulating SARS-CoV-2 in the reverse primer. We did not find evidence for background amplification with pre-COVID-19 samples or recent SARS-CoV-2 evolution decreasing sensitivity. Our recommendation for SARS-CoV-2 diagnostic testing is to select an assay with high sensitivity and that is regionally used, to ease comparability between outcomes.

Original languageEnglish (US)
Pages (from-to)1299-1305
Number of pages7
JournalNature Microbiology
Volume5
Issue number10
DOIs
StatePublished - Oct 1 2020
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Applied Microbiology and Biotechnology
  • Genetics
  • Microbiology (medical)
  • Cell Biology

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