A Novel Multiplex qRT-PCR Assay to Detect SARS-CoV-2 Infection: High Sensitivity and Increased Testing Capacity

Microorganisms

Fiorella Altruda¹  JeanPiero Margaria¹  Miriam Martini¹  Sara Petrillo¹  MariaChiara De Santis¹  Rossana Cavallo²  Paolo Bottino²  Elisa Zanotto²  Giorgia Mandili³  Alessandro Giorgio³  Davide Barberio³  Giovanna Carrà³ 

[1] Department of Molecular Biotechnology and Health Sciences, University of Torino, 10126 Torino, Italy;Department of Public Health and Pediatrics, Microbiology and Virology Unit, Azienda Ospedaliero Universitaria “Città della Salute e della Scienza”, University of Torino, 10126 Torino, Italy;Molecular Biotechnology Center, Via Nizza 52, 10126 Torino, Italy;

关键词: SARS-CoV-2;    COVID-19;    qRT-PCR;    multiplex qRT-PCR;    direct multiplex qRT-PCR;   

DOI  :  10.3390/microorganisms8071064

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown