Laboratory verification of an RT‐PCR assay for SARS‐CoV‐2

Wu, Yingping; Xu, Wei; Zhu, Zhiqiang; Xia, Xiaoping

BACKGROUND: Reverse transcription‐polymerase chain reaction (RT‐PCR) is an extremely common clinical method for detecting pathogens, particularly for emerging infectious diseases such as the new coronavirus disease (COVID‐19). Currently, detection of the RNA from the novel coronavirus SARS‐CoV‐2 is the gold standard for establishing a COVID‐19 diagnosis. This study evaluates the characteristic performance of the analytical system in a clinical laboratory. METHODS: A commercial SARS‐CoV‐2 RNA RT‐PCR Kit used in a clinical laboratory is assessed based on ISO 15189 verification requirements. A multiple real‐time RT‐PCR assay for the RdRP, N, and E genes in SARS‐CoV‐2 is verified. RESULTS: The analytical system exhibits good analytical sensitivity (1000 copies/mL) and specificity (100%); however, the values of 86.7% and 100% for analytical accuracy deserved attention, compared with two other types of methods. Overall, the kit is potentially useful for SARS‐CoV‐2 diagnostic testing and meets the verification requirements. CONCLUSION: Compliance with international standards, such as ISO 15189, is valuable for clinical laboratories and for improving laboratory medicine quality and safety. Normalization is essential for obtaining reliable results from the SARS‐CoV‐2 RNA RT‐PCR assay. This study aims to develop an improved SARS‐CoV‐2 verification framework compared with traditional molecular diagnostic methods, given the urgency of implementing new assays in clinical laboratories.