tetano
Editor, Senior Moderator
Microorganisms
. 2020 Jul 17;8(7):E1064.
doi: 10.3390/microorganisms8071064.
A Novel Multiplex qRT-PCR Assay to Detect SARS-CoV-2 Infection: High Sensitivity and Increased Testing Capacity
Sara Petrillo[SUP] 1 2 [/SUP], Giovanna Carr?[SUP] 2 3 [/SUP], Paolo Bottino[SUP] 4 [/SUP], Elisa Zanotto[SUP] 4 [/SUP], Maria Chiara De Santis[SUP] 1 2 [/SUP], Jean Piero Margaria[SUP] 1 2 [/SUP], Alessandro Giorgio[SUP] 2 [/SUP], Giorgia Mandili[SUP] 2 [/SUP], Miriam Martini[SUP] 1 2 [/SUP], Rossana Cavallo[SUP] 4 [/SUP], Davide Barberio[SUP] 2 [/SUP], Fiorella Altruda[SUP] 1 2 [/SUP]
Affiliations
Abstract
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.
Keywords: COVID-19; SARS-CoV-2; direct multiplex qRT-PCR; multiplex qRT-PCR; qRT-PCR.
. 2020 Jul 17;8(7):E1064.
doi: 10.3390/microorganisms8071064.
A Novel Multiplex qRT-PCR Assay to Detect SARS-CoV-2 Infection: High Sensitivity and Increased Testing Capacity
Sara Petrillo[SUP] 1 2 [/SUP], Giovanna Carr?[SUP] 2 3 [/SUP], Paolo Bottino[SUP] 4 [/SUP], Elisa Zanotto[SUP] 4 [/SUP], Maria Chiara De Santis[SUP] 1 2 [/SUP], Jean Piero Margaria[SUP] 1 2 [/SUP], Alessandro Giorgio[SUP] 2 [/SUP], Giorgia Mandili[SUP] 2 [/SUP], Miriam Martini[SUP] 1 2 [/SUP], Rossana Cavallo[SUP] 4 [/SUP], Davide Barberio[SUP] 2 [/SUP], Fiorella Altruda[SUP] 1 2 [/SUP]
Affiliations
- PMID: 32708870
- DOI: 10.3390/microorganisms8071064
Abstract
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.
Keywords: COVID-19; SARS-CoV-2; direct multiplex qRT-PCR; multiplex qRT-PCR; qRT-PCR.