Establishment of a quantitative RT-PCR detection of SARS-CoV-2 virus

• Published in: European journal of medical research 2021-12, Vol.26 (1), p.147-147, Article 147
• Main Authors: Jiang, Yushen, Zhang, Shanming, Qin, Hong, Meng, Shuai, Deng, Xuyi, Lin, He, Xin, Xiaoliang, Liang, Yuxin, Chen, Bowen, Cui, Yan, Su, YiHeng, Liang, Pei, Zhou, GuangZhi, Hu, Hongbo
• Format: Article
• Language: English
• Subjects: Antiviral agents; Clinical trials; Coronavirus Envelope Proteins - genetics; Coronavirus Nucleocapsid Proteins - genetics; Coronavirus RNA-Dependent RNA Polymerase - genetics; Coronaviruses; COVID-19; COVID-19 - diagnosis; Experiments; Genes; Health aspects; Humans; Infections; Limit of Detection; Phosphoproteins - genetics; Proteins; Quantitative; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA; RNA polymerase; RT-PCR; SARS-CoV-2 - genetics; SARS-CoV-2 - isolation & purification; SARS-CoV-2 virus; Severe acute respiratory syndrome; Severe acute respiratory syndrome coronavirus 2; Thermal cycling; Vaccines; Viral Load - methods
• ISSN: 2047-783X; 0949-2321; 2047-783X
• DOI: 10.1186/s40001-021-00608-5

The outbreak of novel coronavirus disease 2019 (COVID-19) has become a public health emergency of international concern. Quantitative testing of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) virus is demanded in evaluating the efficacy of antiviral drugs and vaccines and RT-PCR can be widely deployed in the clinical assay of viral loads. Here, we developed a quantitative RT-PCR method for SARS-CoV-2 virus detection in this study. RT-PCR kits targeting E (envelope) gene, N (nucleocapsid) gene and RdRP (RNA-dependent RNA polymerase) gene of SARS-CoV-2 from Roche Diagnostics were evaluated and E gene kit was employed for quantitative detection of COVID-19 virus using Cobas Z480. Viral load was calculated according to the standard curve established by series dilution of an E-gene RNA standard provided by Tib-Molbiol (a division of Roche Diagnostics). Assay performance was evaluated. The performance of the assay is acceptable with limit of detection (LOD) below 10E1 copies/μL and lower limit of quantification (LLOQ) as 10E2 copies/μL. A quantitative detection of the COVID-19 virus based on RT-PCR was established.