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Multiplex Real-Time Reverse-Transcription Polymerase Chain Reaction Assays for Diagnostic Testing of Severe Acute Respiratory Syndrome Coronavirus 2 and Seasonal Influenza Viruses: A Challenge of the Phase 3 Pandemic Setting

Authors :
Mancini, Fabiola
Barbanti, Fabrizio
Scaturro, Maria
Fontana, Stefano
Di Martino, Angela
Marsili, Giulia
Puzelli, Simona
Calzoletti, Laura
Facchini, Marzia
Di Mario, Giuseppina
Fabiani, Concetta
Bella, Antonino
Riccardo, Flavia
Pezzotti, Patrizio
Stefanelli, Paola
Rezza, Giovanni
Ciervo, Alessandra
Villa, Laura
Fortini, Daniela
Iacobino, Angelo
Fiore, Stefano
Benedetti, Eleonora
Marchi, Antonella
Venturi, Giulietta
Fortuna, Claudia
Amendola, Antonello
Toma, Luciano
Di Luca, Marco
Severini, Francesco
Source :
The Journal of Infectious Diseases
Publication Year :
2020
Publisher :
Oxford University Press (OUP), 2020.

Abstract

Background Pandemic coronavirus disease 2019 (COVID-19) disease represents a challenge for healthcare structures. The molecular confirmation of samples from infected individuals is crucial and therefore guides public health decision making. Clusters and possibly increased diffuse transmission could occur in the context of the next influenza season. For this reason, a diagnostic test able to discriminate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from influenza viruses is urgently needed. Methods A multiplex real-time reverse-transcription polymerase chain reaction (PCR) assay was assessed using 1 laboratory protocol with different real-time PCR instruments. Overall, 1000 clinical samples (600 from samples SARS-CoV-2–infected patients, 200 samples from influenza-infected patients, and 200 negative samples) were analyzed. Results The assay developed was able to detect and discriminate each virus target and to intercept coinfections. The limit of quantification of each assay ranged between 5 and 10 genomic copy numbers, with a cutoff value of 37.7 and 37.8 for influenza and SARS-CoV-2 viruses, respectively. Only 2 influenza coinfections were detected in COVID-19 samples. Conclusions This study suggests that multiplex assay is a rapid, valid, and accurate method for the detection of SARS-CoV-2 and influenza viruses in clinical samples. The test may be an important diagnostic tool for both diagnostic and surveillance purposes during the seasonal influenza activity period.

Details

Language :
English
ISSN :
15376613 and 00221899
Database :
OpenAIRE
Journal :
The Journal of Infectious Diseases
Accession number :
edsair.doi.dedup.....136f9c238a21ed429101635e0d604fc3
Full Text :
https://doi.org/10.1093/infdis/jiaa658