Open AccessAssessment of Multiplex Digital Droplet RT-PCR as a Diagnostic Tool for SARS-CoV-2 Detection in Nasopharyngeal Swabs and Saliva Samples
Author(s) -
Kévin Cassinari,
Élodie Alessandri-Gradt,
Pascal Chambon,
Françoise Charbonnier,
Ségolène Gracias,
Ludivine Beaussire,
Kévin Alexandre,
Nasrin SarafanVasseur,
Claude Houdayer,
Manuel Etienne,
François Caron,
JeanChristophe Plantier,
Thierry Frébourg
Publication year - 2020
Publication title -
clinical chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.705
H-Index - 218
eISSN - 1530-8561
pISSN - 0009-9147
DOI - 10.1093/clinchem/hvaa323
Subject(s) - saliva , multiplex , digital polymerase chain reaction , multiplex polymerase chain reaction , real time polymerase chain reaction , medicine , covid-19 , reverse transcription polymerase chain reaction , virology , biology , immunology , polymerase chain reaction , bioinformatics , gene , genetics , infectious disease (medical specialty) , messenger rna , disease
Background Reverse transcription-quantitative PCR on nasopharyngeal swabs is currently the reference COVID-19 diagnosis method but exhibits imperfect sensitivity. Methods We developed a multiplex reverse transcription-digital droplet PCR (RT-ddPCR) assay, targeting 6 SARS-CoV-2 genomic regions, and evaluated it on nasopharyngeal swabs and saliva samples collected from 130 COVID-19 positive or negative ambulatory individuals, who presented symptoms suggestive of mild or moderate SARS-CoV2 infection. Results For the nasopharyngeal swab samples, the results obtained using the 6-plex RT-ddPCR and RT-qPCR assays were all concordant. The 6-plex RT-ddPCR assay was more sensitive than RT-qPCR (85% versus 62%) on saliva samples from patients with positive nasopharyngeal swabs. Conclusion Multiplex RT-ddPCR represents an alternative and complementary tool for the diagnosis of COVID-19, in particular to control RT-qPCR ambiguous results. It can also be applied to saliva for repetitive sampling and testing individuals for whom nasopharyngeal swabbing is not possible.
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