Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has rapidly spread worldwide from the beginning of 2020. Quantitative reverse transcription-PCR (RT-qPCR) is, to this day, the preferred methodology for viral RNA detection, even if not without problems. To overcome some of the limitations still existing for the detection and quantification of nucleic acids in various applications, the use of one-step reverse transcription-droplet digital PCR (RT-ddPCR) has been established. The purpose of this study was, then, to evaluate the efficacy of ddPCR for the detection of SARS-CoV-2 RNA in nasopharyngeal swabs, optimizing the detection of low-viral load-burdened samples. Methods The RT-ddPCR workflow was validated for sensitivity, specificity, linearity, reproducibility, and precision using samples from 90 COVID-19-infected patients referred to the Department of Laboratory Medicine of the University Hospital of Udine (Italy). Results The present study shows that RT-ddPCR allows the detection of as low as 10.3 copies of a SARS-COV-2 E-gene per sample with a higher level of accuracy and precision, especially at low concentration. Conclusion During the postpeak phase of the SARS-CoV-2 pandemic, it is essential to rely on a highly robust molecular biology method to identify infected subjects, whether they have symptoms or not, in order to prepare appropriate containment measures.
【초록키워드】 SARS-CoV-2, Efficacy, coronavirus, pandemic, Molecular biology, Infection, Symptom, Italy, Spread, Medicine, digital PCR, nucleic acid, sensitivity, specificity, RT-qPCR, Accuracy, nasopharyngeal swabs, Patient, SARS-CoV-2 RNA, Viral RNA, methodology, university, quantification, E-gene, Concentration, containment measures, acute respiratory syndrome, Precision, problems, limitation, reproducibility, Department, quantitative reverse transcription-PCR, robust, Result, identify, evaluate, subjects, overcome, the SARS-CoV-2, 【제목키워드】 validation, PCR, Digital, SARS-CoV-2 RNA,