Author: Chantal B.F. Vogels; Anderson F. Brito; Anne Louise Wyllie; Joseph R Fauver; Isabel M. Ott; Chaney C. Kalinich; Mary E. Petrone; Marie-Louise Landry; Ellen F. Foxman; Nathan D. Grubaugh
Title: Analytical sensitivity and efficiency comparisons of SARS-COV-2 qRT-PCR assays Document date: 2020_4_1
ID: 6mdimxnk_7
Snippet: Critical evaluations of the designed primer-probe sets used in the primary SARS-CoV-2 qRT-PCR detection assays are necessary to compare findings across studies, and select appropriate assays for in-house testing. The goal of our study was to compare the designed primer-probe sets, not the assays per se , as that would involve many different variables. To do so we used the same ( 1 ) primer-probe concentrations (500 nM of forward and reverse prime.....
Document: Critical evaluations of the designed primer-probe sets used in the primary SARS-CoV-2 qRT-PCR detection assays are necessary to compare findings across studies, and select appropriate assays for in-house testing. The goal of our study was to compare the designed primer-probe sets, not the assays per se , as that would involve many different variables. To do so we used the same ( 1 ) primer-probe concentrations (500 nM of forward and reverse primer, and 250 nM of probe); ( 2 ) PCR reagents (New England Biolabs Luna Universal One-step RT-qPCR kit); and ( 3 ) thermocycler conditions (40 cycles of 10 seconds at 95°C and 20 seconds at 55°C) in all reactions. From our measured PCR amplification efficiencies and analytical sensitivities of detection, most primer-probe sets were comparable, except for the RdRp-SARSr (Charité) set, which had low sensitivity ( Fig. 2 ).
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