Comparative in silico design and validation of GPS™ CoVID‐19 dtec‐RT‐qPCR test

• Published in: Journal of applied microbiology 2021-01, Vol.130 (1), p.2-13
• Main Authors: Martínez‐Murcia, A., Bru, G., Navarro, A., Ros‐Tárraga, P., García‐Sirera, A., Pérez, L.
• Format: Article
• Language: English
• Subjects: Acceptance criteria; Computer Simulation; Coronaviruses; COVID-19; COVID-19 - diagnosis; COVID-19 Nucleic Acid Testing - methods; COVID-19 Nucleic Acid Testing - standards; diagnosis; Diagnostic systems; Editor's Choice; Humans; Laboratories; Original; Pandemics; polymerase chain reaction; Public health; quality control; Real-Time Polymerase Chain Reaction; Reproducibility of Results; RNA-directed DNA polymerase; SARS-CoV-2 - classification; SARS-CoV-2 - genetics; SARS-CoV-2 - isolation & purification; SARS‐CoV‐2; Sensitivity and Specificity; Sequence Alignment; Severe acute respiratory syndrome; Severe acute respiratory syndrome coronavirus 2; Technology transfer
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/jam.14781

Aims Providing a ready‐to‐use reverse transcriptase qPCR (RT‐qPCR) method fully validated to detect the SARS‐CoV‐2 with a higher exclusivity than this shown by early published RT‐qPCR designs. Methods and Results The specificity of the GPS™ CoVID‐19 dtec‐RT‐qPCR test by analysis of sequence alignments was approached and compared with other RT‐qPCR designs. The GPS™ CoVID‐19 dtec‐RT‐qPCR test was validated following criteria of UNE/EN ISO 17025:2005 and ISO/IEC 15189:2012. Diagnostic validation was achieved by two independent reference laboratories, the Instituto de Salud Carlos III, (Madrid, Spain), the Public Health England (Colindale, London, UK), and received the label CE‐IVD. The GPS design showed the highest exclusivity and passed all parameters of validation with strict acceptance criteria. Results from reference laboratories 100% correlated with these obtained by using reference methods and showed 100% of diagnostic sensitivity and specificity. Conclusions The CE‐IVD GPS™ CoVID‐19 dtec‐RT‐qPCR test, available worldwide with full analytical and diagnostic validation, is the more exclusive for SARS‐CoV‐2 by far. Significance and Impact of the Study Considering the CoVID‐19 pandemic status, the exclusivity of RT‐qPCR tests is crucial to avoid false positives due to related coronaviruses. This work provides of a highly specific and validated RT‐qPCR method for detection of SARS‐CoV‐2, which represents a case of efficient transfer of technology successfully used since the pandemic was declared.