Development of RT-LAMP and real-time RT-PCR assays for the rapid detection of the new duck Tembusu-like BYD virus

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• 作者：Tao Jiang (1)
  Juan Liu (1) (2)
  Yong-Qiang Deng (1)
  Jing-Liang Su (3)
  Li-Juan Xu (1)
  Zhi-Hui Liu (1) (4)
  Xiao-Feng Li (1)
  Xue-Dong Yu (1)
  Shun-Ya Zhu (1)
  George Fu Gao (5)
  E-De Qin (1)
  Cheng-Feng Qin (1)
  
• 刊名：Archives of Virology
• 出版年：2012
• 出版时间：December 2012
• 年：2012
• 卷：157
• 期：12
• 页码：2273-2280
• 全文大小：436KB
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• 作者单位：Tao Jiang (1)
  Juan Liu (1) (2)
  Yong-Qiang Deng (1)
  Jing-Liang Su (3)
  Li-Juan Xu (1)
  Zhi-Hui Liu (1) (4)
  Xiao-Feng Li (1)
  Xue-Dong Yu (1)
  Shun-Ya Zhu (1)
  George Fu Gao (5)
  E-De Qin (1)
  Cheng-Feng Qin (1)
  
  1. State Key Laboratory of Pathogen and Biosecurity, Department of Virology, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, 100071, Beijing, China
  2. Department of Transfusion Medicine, PLA Air Force General Hospital, 100142, Beijing, China
  3. Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, China
  4. College of Veterinary Medicine, Xinjiang Agricultural University, 830052, Urumqi, China
  5. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China
  
• ISSN：1432-8798

文摘

A new duck Tembusu virus (TMUV), also known as BYD virus, has been identified as the causative agent for the emerging duck egg-drop syndrome in mainland China. The rapid spread and wide distribution of the new TMUV in mainland China result in heavy loss to the poultry industry and pose great threats to public health. Rapid and sensitive detection methods are critical for prevention and control of TMUV infections. In this study, a reverse-transcription loop-mediated isothermal amplification assay (RT-LAMP) and an SYBR Green-I–based real-time RT-PCR assay specific for the duck TMUV were developed and validated with laboratory and field samples, respectively. The detection limits were 1?×?10? and 1?×?10? PFU per reaction for the RT-LAMP assay and real-time RT-PCR assay, respectively. The specificities were analyzed with other related members of the genus Flavivirus, and no cross-reaction was observed. Furthermore, both assays were evaluated with field samples, and they exhibited high sensitivity and specificity. In addition, the real-time RT-PCR assay worked well in viral load analysis, which revealed that the spleen may be the primary target for the replication of new duck TMUV in ducks. The TMUV-specific RT-LAMP and real-time RT-PCR assays will provide useful tools for the diagnosis and epidemiological surveillance of TMUV infection.