Messenger RNA

Internal medicine

Disease

Coronavirus disease 2019 (COVID-19)

Saliva

Infectious disease (medical specialty)

Bioinformatics

Biology

Gene

Virology

Immunology

Digital polymerase chain reaction

Multiplex

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)

Reverse transcription polymerase chain reaction

Real-time polymerase chain reaction

Polymerase chain reaction

Genetics

Medicine

Multiplex polymerase chain reaction

Clinical Chemistry

Reverse transcription-quantitative PCR on nasopharyngeal swabs is currently the reference COVID-19 diagnosis method but exhibits imperfect sensitivity.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.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.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.

Assessment of Multiplex Digital Droplet RT-PCR as a Diagnostic Tool for SARS-CoV-2 Detection in Nasopharyngeal Swabs and Saliva Samples