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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 Kai-Wang To.
Helen Shuk-Ying Chan.
Sally Cheuk-Ying Wong.
Kit-Hang Leung.
Agnes Yim-Fong Fung.
Anthony Chin-Ki Ng.
Zijiao Zou.
Anthony Raymond Tam.
Tom Wai-Hin Chung.
Kwok-Hung Chan.
Ivan Fan-Ngai Hung.
Vincent Chi-Chung Cheng.
Owen Tak-Yin Tsang.
Stephen Kwok Wing Tsui.
Kwok-Yung Yuen.
Acceso Abierto
Atribución-NoComercial-SinDerivadas
10.3390/ijms21072574
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
2020
Artículo
https://www.mdpi.com/1422-0067/21/7/2574/pdf
Inglés
VIRUS RESPIRATORIOS
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