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Virologica Sinica, 31 (1) : 85-88, 2016
Two-tube multiplex real-time reverse transcription PCR to detect six human coronaviruses
1. Key Laboratory of Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
2. Children’s Hospital of Fudan University, Shanghai 200032,
 Correspondence: tanwj28@163.com
(230.51KB)  (4191.96KB)  
In this study, the authors developed a two-tube multiplex real-time RT-PCR assay for sensitive and specific detection of all known human coronaviruses. Its ability to monitor HKU1 replication in cultures of human airway epithelial cells, to quantitatively measure viral RNA in monkeys experimentally infected with MERS, and to detect MERS in a human patient was demonstrated. In addition, the assay was used to assess disease burden and epidemiology of coronaviruses among hospitalized patients with acute respiratory infection. The results indicate that the assay is quantitative, rapid, sensitive, specific, high-throughput, and able to detect co-infection. Finally, the assay requires significantly less sample than monoplex real-time RT-PCR.
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Received: 23 Sep 2015  Accepted: 6 Jan 2016  Published online: 25 Jan 2016
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