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Microorganisms
Volume 8
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10.3390/microorganisms8071064
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Communication

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

by
Sara Petrillo
1,2,*,†,
Giovanna Carrà
2,3,*,†,
Paolo Bottino
4,
Elisa Zanotto
4,
Maria Chiara De Santis
1,2,
Jean Piero Margaria
1,2,
Alessandro Giorgio
2,
Giorgia Mandili
2,
Miriam Martini
1,2,
Rossana Cavallo
4,
Davide Barberio
2,‡ and
Fiorella Altruda
1,2,‡
1
Department of Molecular Biotechnology and Health Sciences, University of Torino, 10126 Torino, Italy
2
Molecular Biotechnology Center, Via Nizza 52, 10126 Torino, Italy
3
Department of Clinical and Biological Sciences, University of Torino, 10043 Orbassano, Italy
4
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
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
‡
These authors contributed equally to this work.
Microorganisms 2020, 8(7), 1064; https://doi.org/10.3390/microorganisms8071064
Submission received: 23 June 2020 / Revised: 13 July 2020 / Accepted: 14 July 2020 / Published: 17 July 2020
(This article belongs to the Special Issue COVID-19: Focusing on Epidemiologic, Virologic, and Clinical Studies)
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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: SARS-CoV-2; COVID-19; qRT-PCR; multiplex qRT-PCR; direct multiplex qRT-PCR SARS-CoV-2; COVID-19; qRT-PCR; multiplex qRT-PCR; direct multiplex qRT-PCR

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MDPI and ACS Style

Petrillo, S.; Carrà, G.; Bottino, P.; Zanotto, E.; De Santis, M.C.; Margaria, J.P.; Giorgio, A.; Mandili, G.; Martini, M.; Cavallo, R.; et al. A Novel Multiplex qRT-PCR Assay to Detect SARS-CoV-2 Infection: High Sensitivity and Increased Testing Capacity. Microorganisms 2020, 8, 1064. https://doi.org/10.3390/microorganisms8071064

AMA Style

Petrillo S, Carrà G, Bottino P, Zanotto E, De Santis MC, Margaria JP, Giorgio A, Mandili G, Martini M, Cavallo R, et al. A Novel Multiplex qRT-PCR Assay to Detect SARS-CoV-2 Infection: High Sensitivity and Increased Testing Capacity. Microorganisms. 2020; 8(7):1064. https://doi.org/10.3390/microorganisms8071064

Chicago/Turabian Style

Petrillo, Sara, Giovanna Carrà, Paolo Bottino, Elisa Zanotto, Maria Chiara De Santis, Jean Piero Margaria, Alessandro Giorgio, Giorgia Mandili, Miriam Martini, Rossana Cavallo, and et al. 2020. "A Novel Multiplex qRT-PCR Assay to Detect SARS-CoV-2 Infection: High Sensitivity and Increased Testing Capacity" Microorganisms 8, no. 7: 1064. https://doi.org/10.3390/microorganisms8071064

APA Style

Petrillo, S., Carrà, G., Bottino, P., Zanotto, E., De Santis, M. C., Margaria, J. P., Giorgio, A., Mandili, G., Martini, M., Cavallo, R., Barberio, D., & Altruda, F. (2020). A Novel Multiplex qRT-PCR Assay to Detect SARS-CoV-2 Infection: High Sensitivity and Increased Testing Capacity. Microorganisms, 8(7), 1064. https://doi.org/10.3390/microorganisms8071064

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