tetano
Editor, Senior Moderator
Viruses
. 2020 Jun 25;12(6):E686.
doi: 10.3390/v12060686.
Development and Evaluation of a Duo SARS-CoV-2 RT-qPCR Assay Combining Two Assays Approved by the World Health Organization Targeting the Envelope and the RNA-Dependant RNA Polymerase (RdRp) Coding Regions
Laura Pezzi[SUP] 1 2 [/SUP], Remi N Charrel[SUP] 2 [/SUP], Laetitia Ninove[SUP] 2 [/SUP], Antoine Nougairede[SUP] 2 [/SUP], Gregory Molle[SUP] 2 [/SUP], Bruno Coutard[SUP] 2 [/SUP], Guillaume Durand[SUP] 2 3 [/SUP], Isabelle Leparc-Goffart[SUP] 2 3 [/SUP], Xavier de Lamballerie[SUP] 2 [/SUP], Laurence Thirion[SUP] 2 [/SUP]
Affiliations
Abstract
The recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide has highlighted the importance of reliable and rapid diagnostic testing to prevent and control virus circulation. Dozens of monoplex in-house RT-qPCR assays are already available; however, the development of dual-target assays is suited to avoid false-negative results caused by polymorphisms or point mutations, that can compromise the accuracy of diagnostic and screening tests. In this study, two mono-target assays recommended by WHO (E-Sarbeco (enveloppe gene, Charite University, Berlin, Germany) and RdRp-IP4 (RdRp, Institut Pasteur, Paris, France)) were selected and combined in a unique robust test; the resulting duo SARS-CoV-2 RT-qPCR assay was compared to the two parental monoplex tests. The duo SARS-CoV-2 assay performed equally, or better, in terms of sensitivity, specificity, linearity and signal intensity. We demonstrated that combining two single systems into a dual-target assay (with or without an MS2-based internal control) did not impair performances, providing a potent tool adapted for routine molecular diagnosis in clinical microbiology laboratories.
Keywords: COVID-19; TaqMan; coronavirus; diagnosis; diagnostics; emerging; molecular; outbreak; preparedness; real-time PCR; respiratory; response.
. 2020 Jun 25;12(6):E686.
doi: 10.3390/v12060686.
Development and Evaluation of a Duo SARS-CoV-2 RT-qPCR Assay Combining Two Assays Approved by the World Health Organization Targeting the Envelope and the RNA-Dependant RNA Polymerase (RdRp) Coding Regions
Laura Pezzi[SUP] 1 2 [/SUP], Remi N Charrel[SUP] 2 [/SUP], Laetitia Ninove[SUP] 2 [/SUP], Antoine Nougairede[SUP] 2 [/SUP], Gregory Molle[SUP] 2 [/SUP], Bruno Coutard[SUP] 2 [/SUP], Guillaume Durand[SUP] 2 3 [/SUP], Isabelle Leparc-Goffart[SUP] 2 3 [/SUP], Xavier de Lamballerie[SUP] 2 [/SUP], Laurence Thirion[SUP] 2 [/SUP]
Affiliations
- PMID: 32630601
- DOI: 10.3390/v12060686
Abstract
The recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide has highlighted the importance of reliable and rapid diagnostic testing to prevent and control virus circulation. Dozens of monoplex in-house RT-qPCR assays are already available; however, the development of dual-target assays is suited to avoid false-negative results caused by polymorphisms or point mutations, that can compromise the accuracy of diagnostic and screening tests. In this study, two mono-target assays recommended by WHO (E-Sarbeco (enveloppe gene, Charite University, Berlin, Germany) and RdRp-IP4 (RdRp, Institut Pasteur, Paris, France)) were selected and combined in a unique robust test; the resulting duo SARS-CoV-2 RT-qPCR assay was compared to the two parental monoplex tests. The duo SARS-CoV-2 assay performed equally, or better, in terms of sensitivity, specificity, linearity and signal intensity. We demonstrated that combining two single systems into a dual-target assay (with or without an MS2-based internal control) did not impair performances, providing a potent tool adapted for routine molecular diagnosis in clinical microbiology laboratories.
Keywords: COVID-19; TaqMan; coronavirus; diagnosis; diagnostics; emerging; molecular; outbreak; preparedness; real-time PCR; respiratory; response.