pone.0243333.g001.tif (280.85 kB)

Performance and validation of the Biomark-HD SARS-CoV-2 qPCR assay.

figure

posted on 2021-04-14, 17:55 authored by Julien Fassy, Caroline Lacoux, Sylvie Leroy, Latifa Noussair, Sylvain Hubac, Aurélien Degoutte, Georges Vassaux, Vianney Leclercq, David Rouquié, Charles-Hugo Marquette, Martin Rottman, Patrick Touron, Antoinette Lemoine, Jean-Louis Herrmann, Pascal Barbry, Jean-Louis Nahon, Laure-Emmanuelle Zaragosi, Bernard Mari

(A) Titration of the diluted (N) nucleocapside spike-in transcript. A serial dilution of the synthetic N transcript from 2.10⁴ to 2 copies was performed and processed through the Biomark-HD protocol. Correlation and amplification curves of detected Cq values according to synthetic N transcript copy number obtained with N1 primers/probe are shown. (B) Amplification curves showing the range of detection of a SARS-CoV-2 positive clinical sample serial dilution with N1 and E primers/probe. (C) Typical amplification curves showing the combination of two primers/probe sets on 3 SARS-CoV-2 positive clinical samples. Mix #1: E, N1, human RNP. Mix #2: E, N, ORF1ab and human RNP. (D) Validation of the Biomark-HD protocol on a cohort of 18 biopsies from positive patients. The correlation of the Cq values obtained for the N, E and the ORF1ab genes are presented. The data presented are representative of at least two independent experiments performed in quadruplicate.