Accuracy of Real-Time Polymerase Chain Reaction in COVID-19 Patients

Autor: Merlin Jayalal Lawrence Panchali, Hyeon Jeong Oh, You Mi Lee, Choon-Mee Kim, Misbah Tariq, Jun-Won Seo, Da Young Kim, Na Ra Yun, Dong-Min Kim
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Microbiology Spectrum, Vol 10, Iss 1 (2022)
Druh dokumentu: article
ISSN: 2165-0497
DOI: 10.1128/spectrum.00591-21
Popis: ABSTRACT Coronavirus disease 2019 (COVID-19) is a mild to severe respiratory illness caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The diagnostic accuracy of the Centers for Disease Control and Prevention (CDC)- or World Health Organization (WHO)-recommended real-time PCR (RT-qPCR) primers in clinical practice remains unproven. We conducted a prospective study on the accuracy of RT-qPCR using an in-house–designed primer set (iNP) targeting the nucleocapsid protein as well as various recommended and commercial primers. The accuracy was assessed by culturing or seroconversion. We enrolled 12 confirmed COVID-19 patients with a total of 590 clinical samples. When a cutoff value of the cycle threshold (Ct) was set to 35, RT-qPCRs with WHO RdRp primers and CDC N1, N2, and N3 primers showed sensitivity of 42.1% to 63.2% and specificity of 90.5% to 100% in sputum, and sensitivity of 65.2% to 69.6% and specificity of 65.2% to 69.6% in nasopharyngeal samples. The sensitivity and specificity of iNP RT-qPCR in sputum and nasopharyngeal samples were 94.8%/100% and 69.6%/100%, respectively. Sputum testing had the highest sensitivity, followed by nasopharyngeal testing (P = 0.0193); self-collected saliva samples yielded better characteristics than oropharyngeal samples (P = 0.0032). Our results suggest that iNP RT-qPCR has better sensitivity and specificity than RT-PCR with WHO (P
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