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ddPCR: a more accurate tool for SARS-CoV-2 detection in low viral load specimens.
Suo, Tao; Liu, Xinjin; Feng, Jiangpeng; Guo, Ming; Hu, Wenjia; Guo, Dong; Ullah, Hafiz; Yang, Yang; Zhang, Qiuhan; Wang, Xin; Sajid, Muhanmmad; Huang, Zhixiang; Deng, Liping; Chen, Tielong; Liu, Fang; Xu, Ke; Liu, Yuan; Zhang, Qi; Liu, Yingle; Xiong, Yong; Chen, Guozhong; Lan, Ke; Chen, Yu.
  • Suo T; State Key Laboratory of Virology, Renmin Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Liu X; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Feng J; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Guo M; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Hu W; Department of Infectious Disease, Zhongnan Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Guo D; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Ullah H; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Yang Y; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Zhang Q; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Wang X; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Sajid M; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Huang Z; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Deng L; Department of Infectious Disease, Zhongnan Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Chen T; Department of Infectious Disease, Zhongnan Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Liu F; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Xu K; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Liu Y; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Zhang Q; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Liu Y; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Xiong Y; Department of Infectious Disease, Zhongnan Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Chen G; State Key Laboratory of Virology, Renmin Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Lan K; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Chen Y; Frontier Science Center for Immunology and Metabolism, Wuhan University, Wuhan, People's Republic of China.
Emerg Microbes Infect ; 9(1): 1259-1268, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: covidwho-342833
ABSTRACT
Quantitative real time PCR (RT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. However, due to the low viral load specimens and the limitations of RT-PCR, significant numbers of false negative reports are inevitable, which results in failure to timely diagnose, cut off transmission, and assess discharge criteria. To improve this situation, an optimized droplet digital PCR (ddPCR) was used for detection of SARS-CoV-2, which showed that the limit of detection of ddPCR is significantly lower than that of RT-PCR. We further explored the feasibility of ddPCR to detect SARS-CoV-2 RNA from 77 patients, and compared with RT-PCR in terms of the diagnostic accuracy based on the results of follow-up survey. 26 patients of COVID-19 with negative RT-PCR reports were reported as positive by ddPCR. The sensitivity, specificity, PPV, NPV, negative likelihood ratio (NLR) and accuracy were improved from 40% (95% CI 27-55%), 100% (95% CI 54-100%), 100%, 16% (95% CI 13-19%), 0.6 (95% CI 0.48-0.75) and 47% (95% CI 33-60%) for RT-PCR to 94% (95% CI 83-99%), 100% (95% CI 48-100%), 100%, 63% (95% CI 36-83%), 0.06 (95% CI 0.02-0.18), and 95% (95% CI 84-99%) for ddPCR, respectively. Moreover, 6/14 (42.9%) convalescents were detected as positive by ddPCR at 5-12 days post discharge. Overall, ddPCR shows superiority for clinical diagnosis of SARS-CoV-2 to reduce the false negative reports, which could be a powerful complement to the RT-PCR.
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Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: MEDLINE Assunto principal: Pneumonia Viral / Infecções por Coronavirus / Reação em Cadeia da Polimerase em Tempo Real / Betacoronavirus Tipo de estudo: Estudo de coorte / Estudo diagnóstico / Estudo observacional / Estudo prognóstico Tópicos: Covid persistente Limite: Humanos Idioma: Inglês Revista: Emerg Microbes Infect Ano de publicação: 2020 Tipo de documento: Artigo

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Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: MEDLINE Assunto principal: Pneumonia Viral / Infecções por Coronavirus / Reação em Cadeia da Polimerase em Tempo Real / Betacoronavirus Tipo de estudo: Estudo de coorte / Estudo diagnóstico / Estudo observacional / Estudo prognóstico Tópicos: Covid persistente Limite: Humanos Idioma: Inglês Revista: Emerg Microbes Infect Ano de publicação: 2020 Tipo de documento: Artigo