Rapid and sensitive screening of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is essential to limit the spread of the global pandemic we are facing. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is currently used for the clinical diagnosis of SARS-CoV-2 infection using nasopharyngeal swabs, tracheal aspirates, or bronchoalveolar lavage (BAL) samples. Despite the high sensitivity of the qRT-PCR method, false negative outcomes might occur, especially in patients with a low viral load. Here, we developed a multiplex qRT-PCR methodology for the simultaneous detection of SARS-CoV-2 genome ( N gene) and of the human RNAse P gene as internal control. We found that multiplex qRT-PCR was effective in detecting SARS-Cov-2 infection in human specimens with 100% sensitivity. Notably, patients with few copies of SARS-CoV-2 RNA (<5 copies/reaction) were successfully detected by the novel multiplex qRT-PCR method. Finally, we assessed the efficacy of multiplex qRT-PCR on human nasopharyngeal swabs without RNA extraction. Collectively, our results provide evidence of a novel and reliable tool for SARS-CoV-2 RNA detection in human specimens, which allows the testing capacity to be expanded and the RNA extraction step to be bypassed.
【저자키워드】 COVID-19, SARS-CoV-2, qRT-PCR, multiplex qRT-PCR, direct multiplex qRT-PCR, 【초록키워드】 Efficacy, coronavirus, SARS-COV-2 infection, Infection, outcome, global pandemic, Spread, RNA extraction, Nasopharyngeal swab, BAL, sensitivity, Internal control, Viral load, nasopharyngeal swabs, SARS-CoV-2 genome, Patient, Rapid, SARS-CoV-2 RNA, methodology, N gene, multiplex, Clinical diagnosis, specimens, Evidence, Bronchoalveolar lavage, SARS-CoV-2 RNA detection, acute respiratory syndrome, Tracheal aspirates, specimen, effective, limit, occur, bypassed, 【제목키워드】 Testing, Capacity, novel, Increased,