Improved Molecular Diagnosis of COVID-19 by the Novel, Highly Sensitive and Specific COVID-19-RdRp/Hel Real-Time Reverse Transcription-PCR Assay Validated
| العنوان: | Improved Molecular Diagnosis of COVID-19 by the Novel, Highly Sensitive and Specific COVID-19-RdRp/Hel Real-Time Reverse Transcription-PCR Assay Validated |
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| المؤلفون: | Jasper Fuk-Woo, Chan, Cyril Chik-Yan, Yip, Kelvin Kai-Wang, To, Tommy Hing-Cheung, Tang, Sally Cheuk-Ying, Wong, Kit-Hang, Leung, Agnes Yim-Fong, Fung, Anthony Chin-Ki, Ng, Zijiao, Zou, Hoi-Wah, Tsoi, Garnet Kwan-Yue, Choi, Anthony Raymond, Tam, Vincent Chi-Chung, Cheng, Kwok-Hung, Chan, Owen Tak-Yin, Tsang, Kwok-Yung, Yuen |
| المصدر: | Journal of Clinical Microbiology |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Adult, Male, Wuhan, China, COVID-19 Vaccines, viruses, Pneumonia, Viral, coronavirus, diagnostic, virus, In Vitro Techniques, Sensitivity and Specificity, Betacoronavirus, Viral Proteins, COVID-19 Testing, Virology, Chlorocebus aethiops, Animals, Humans, Pandemics, Vero Cells, Aged, SARS, Clinical Laboratory Techniques, Reverse Transcriptase Polymerase Chain Reaction, SARS-CoV-2, emerging, virus diseases, COVID-19, Middle Aged, PCR, Molecular Diagnostic Techniques, Female, Coronavirus Infections |
| الوصف: | On 31 December 2019, the World Health Organization was informed of a cluster of cases of pneumonia of unknown etiology in Wuhan, China. Subsequent investigations identified a novel coronavirus, now named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), from the affected patients. Highly sensitive and specific laboratory diagnostics are important for controlling the rapidly evolving SARS-CoV-2-associated coronavirus disease 2019 (COVID-19) epidemic. In this study, we developed and compared the performance of three novel real-time reverse transcription-PCR (RT-PCR) assays targeting the RNA-dependent RNA polymerase (RdRp)/helicase (Hel), spike (S), and nucleocapsid (N) genes of SARS-CoV-2 with that of the reported RdRp-P2 assay, which is used in >30 European laboratories. On 31 December 2019, the World Health Organization was informed of a cluster of cases of pneumonia of unknown etiology in Wuhan, China. Subsequent investigations identified a novel coronavirus, now named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), from the affected patients. Highly sensitive and specific laboratory diagnostics are important for controlling the rapidly evolving SARS-CoV-2-associated coronavirus disease 2019 (COVID-19) epidemic. In this study, we developed and compared the performance of three novel real-time reverse transcription-PCR (RT-PCR) assays targeting the RNA-dependent RNA polymerase (RdRp)/helicase (Hel), spike (S), and nucleocapsid (N) genes of SARS-CoV-2 with that of the reported RdRp-P2 assay, which is used in >30 European laboratories. Among the three novel assays, the COVID-19-RdRp/Hel assay had the lowest limit of detection in vitro (1.8 50% tissue culture infective doses [TCID50]/ml with genomic RNA and 11.2 RNA copies/reaction with in vitro RNA transcripts). Among 273 specimens from 15 patients with laboratory-confirmed COVID-19 in Hong Kong, 77 (28.2%) were positive by both the COVID-19-RdRp/Hel and RdRp-P2 assays. The COVID-19-RdRp/Hel assay was positive for an additional 42 RdRp-P2-negative specimens (119/273 [43.6%] versus 77/273 [28.2%]; P < 0.001), including 29/120 (24.2%) respiratory tract specimens and 13/153 (8.5%) non-respiratory tract specimens. The mean viral load of these specimens was 3.21 × 104 RNA copies/ml (range, 2.21 × 102 to 4.71 × 105 RNA copies/ml). The COVID-19-RdRp/Hel assay did not cross-react with other human-pathogenic coronaviruses and respiratory pathogens in cell culture and clinical specimens, whereas the RdRp-P2 assay cross-reacted with SARS-CoV in cell culture. The highly sensitive and specific COVID-19-RdRp/Hel assay may help to improve the laboratory diagnosis of COVID-19. |
| تدمد: | 1098-660X |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=pmid________::52f198312e5b7b64a29041c147a3b337Test https://pubmed.ncbi.nlm.nih.gov/32132196Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.pmid..........52f198312e5b7b64a29041c147a3b337 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 1098660X |
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