Abstract
The emergence and spread of SARS-CoV-2 has led to a compelling request for accurate diagnostic tests. The aim of this study was assessing the performance of a real-time RT-qPCR (rt RT-qPCR) assay and of a droplet digital RT-PCR (dd RT-PCR) targeting the nsp14 genome region for the detection of SARS-CoV-2 in nasopharyngeal swabs. A total of 258 nasopharyngeal swabs were analyzed with the nsp14 assays and, for comparison, with a reference assay targeting the RdRp and E genes. Conflicting results were further investigated by two additional protocols, the Centers for Disease Control and Prevention (CDC) real-time targeting N1/N2, and a nested RT-PCR for the spike region. Agreement of results was achieved on 226 samples (156 positive and 70 negative), 8 samples were positive in the reference assay and in the nsp14 rt RT-qPCR but negative with the dd RT-PCR, and 24 samples provided different combinations of results with the three assays. Sensitivity, specificity and accuracy (95 %C.I.) of the nsp14 assays were: 100.0 % (97.4-100.0), 98.7 % (92.1-100.0), and 99.6 % (97.5-100.0) for the rt RT-qPCR; 92.4 % (87.4-95.6), 100.0 % (94.2-100.0), and 94.7 % (91.1-97.0) for the dd RT-PCR. The results of the study support the use of the nsp14 real-time RT-qPCR and ddPCR for the detection of SARS-CoV-2 in nasopharyngeal swabs.
Keywords: Droplet digital RT-PCR; Polymerase chain reaction; Real-time RT-qPCR; SARS-CoV-2; Swab; nsp14.
【저자키워드】 SARS-CoV-2, polymerase chain reaction, nsp14, Swab, Droplet digital RT-PCR, Real-time RT-qPCR, 【초록키워드】 Genome, droplet, prevention, RT-PCR, Spread, Nasopharyngeal swab, polymerase chain reaction, specificity, RT-qPCR, Accuracy, CDC, nasopharyngeal swabs, diagnostic tests, qPCR, nsp14, disease control, Control, RdRP, Swab, agreement, Combination, protocols, Chain Reaction, Support, request for, center, positive, e genes, analyzed, assays, investigated, provided, 【제목키워드】 SARS-CoV-2 detection, RT-qPCR, Characteristics, nasopharyngeal, performance, RT-PCR assay, exonuclease, ORF1b,