Please use this identifier to cite or link to this item:
Development of a Novel, Genome Subtraction-Derived, SARS-CoV-2-Specific COVID-19-nsp2 Real-Time RT-PCR Assay and Its Evaluation Using Clinical Specimens
Cyril Chik-Yan Yip
Chi-Chun Ho
Jasper Fuk-Woo Chan
Kelvin To
Shuk Ying Chan
Cheuk Yiu WONG
Raymond Tam
Tom Wai-Hin Chung
Kwok Hung Chan
Ivan Fan Ngai Hung
Owen Tak-Yin Tsang
Stephen Kwok-Wing Tsui
Kwok_yung Yuen
Acceso Abierto
Real-time RT-PCR
Genome subtraction
Clinical evaluation
COVID-19-nsp2 assay
The pandemic novel coronavirus infection, Coronavirus Disease 2019 (COVID-19), has affected at least 190 countries or territories, with 465,915 confirmed cases and 21,031 deaths. In a containment-based strategy, rapid, sensitive and specific testing is important in epidemiological control and clinical management. Using 96 SARS-CoV-2 and 104 non-SARS-CoV-2 coronavirus genomes and our in-house program, GolayMetaMiner, four specific regions longer than 50 nucleotides in the SARS-CoV-2 genome were identified. Primers were designed to target the longest and previously untargeted nsp2 region and optimized as a probe-free real-time reverse transcription-polymerase chain reaction (RT-PCR) assay. The new COVID-19-nsp2 assay had a limit of detection (LOD) of 1.8 TCID50/mL and did not amplify other human-pathogenic coronaviruses and respiratory viruses. Assay reproducibility in terms of cycle threshold (Cp) values was satisfactory, with the total imprecision (% CV) values well below 5%. Evaluation of the new assay using 59 clinical specimens from 14 confirmed cases showed 100% concordance with our previously developed COVID-19-RdRp/Hel reference assay. A rapid, sensitive, SARS-CoV-2-specific real-time RT-PCR assay, COVID-19-nsp2, was developed.
International Journal of Molecular Sciences
Epidemia COVID-19
Versión publicada
publishedVersion - Versión publicada
Appears in Collections:Artículos científicos

Upload archives