The recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide has highlighted the importance of reliable and rapid diagnostic testing to prevent and control virus circulation. Dozens of monoplex in-house RT-qPCR assays are already available; however, the development of dual-target assays is suited to avoid false-negative results caused by polymorphisms or point mutations, that can compromise the accuracy of diagnostic and screening tests. In this study, two mono-target assays recommended by WHO (E-Sarbeco (enveloppe gene, Charite University, Berlin, Germany) and RdRp-IP4 (RdRp, Institut Pasteur, Paris, France)) were selected and combined in a unique robust test; the resulting duo SARS-CoV-2 RT-qPCR assay was compared to the two parental monoplex tests. The duo SARS-CoV-2 assay performed equally, or better, in terms of sensitivity, specificity, linearity and signal intensity. We demonstrated that combining two single systems into a dual-target assay (with or without an MS2-based internal control) did not impair performances, providing a potent tool adapted for routine molecular diagnosis in clinical microbiology laboratories.
【저자키워드】 COVID-19, coronavirus, Diagnosis, diagnostics, Real-time PCR, outbreak, response, emerging, molecular, respiratory, preparedness, TaqMan, 【초록키워드】 SARS-CoV-2, mutations, polymorphism, diagnostic, virus, sensitivity, specificity, RT-qPCR, Molecular diagnosis, Accuracy, Internal control, Germany, RdRP, WHO, circulation, university, Berlin, false-negative, RT-qPCR assay, screening tests, intensity, acute respiratory syndrome, Paris, Prevent, robust, selected, resulting, performed, caused, unique, demonstrated, parental, impair, 【제목키워드】 SARS-CoV-2, RNA, RT-qPCR, Region, RdRP, envelope, development, World Health Organization, targeting,