Assessment of Multiplex Digital Droplet RT-PCR as a Diagnostic Tool for SARS-CoV-2 Detection in Nasopharyngeal Swabs and Saliva Samples
| Autor: | Thierry Frebourg, Kévin Cassinari, Ségolène Gracias, Kevin Alexandre, Jean-Christophe Plantier, Ludivine Beaussire, Manuel Etienne, François Caron, Elodie Alessandri-Gradt, Nasrin Sarafan-Vasseur, Pascal Chambon, Françoise Charbonnier, Claude Houdayer |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
2019-20 coronavirus outbreak Saliva Coronavirus disease 2019 (COVID-19) Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Clinical Biochemistry Article Specimen Handling 03 medical and health sciences 0302 clinical medicine stomatognathic system Limit of Detection Nasopharynx Humans Medicine Sampling (medicine) Multiplex 030212 general & internal medicine Biochemistry medical saliva SARS-CoV-2 Reverse Transcriptase Polymerase Chain Reaction business.industry Biochemistry (medical) COVID-19 Reproducibility of Results RT-ddPCR sensitivity nasopharyngeal swab Virology 030104 developmental biology Real-time polymerase chain reaction COVID-19 Nucleic Acid Testing RNA Viral business |
| Zdroj: | Clinical Chemistry |
| ISSN: | 1530-8561 0009-9147 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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