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Yip, Cyril Chik-Yan; Ho, Chi-Chun; Chan, Jasper Fuk-Woo; To, Kelvin Kai-Wang; Chan, Helen Shuk-Ying; Wong, Sally Cheuk-Ying; Leung, Kit-Hang; Fung, Agnes Yim-Fong; Ng, Anthony Chin-Ki; Zou, Zijiao; Tam, Anthony Raymond; Chung, Tom Wai-Hin; Chan, Kwok-Hung; Hung, Ivan Fan-Ngai; Cheng, Vincent Chi-Chung; Tsang, Owen Tak-Yin; Tsui, Stephen Kwok Wing; Yuen, Kwok-Yung
International journal of molecular sciences, 04/2020, Letnik: 21, Številka: 7Journal Article
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 TCID /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.
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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in: SICRIS
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