Establishment of Pyrosequencing Technology for Detecting Viral Hemorrhagic Septicemia Virus (VHSV)  

作　　者：Yin Weili Liu Yao Huang Wei Zhang Sihua Yue Zhiqin Sun Tao 

机构地区：[1]Yantai Entry-Exit Inspection and Quarantine Bureau [2]Rongcheng Entry-Exit Inspection and Quarantine Bureau [3]Wuhan Agricultural Testing Center [4]Wuhan Animal Disease Control and Prevention Center [5]Technical Center of Shandong Entry-Exit Inspection and Quarantine Bureau

出　　处：《Animal Husbandry and Feed Science》2017年第6期407-410,共4页动物与饲料科学（英文版）

基　　金：Supported by the Twelfth Five-Year Support Project of the Ministry of Science and Technology(2013BAD12B02);Science and Technology Project of State General Administration of the People’s Republic of China for Quality Supervision and Inspection and Quarantine(2015IK195)

摘　　要：[Objective] The paper was to establish pyrosequencing methods for detecting viral hemorrhagic septicemia virus （VHSV）. [ Method ] One pair of PCR primers and one pyrosequencing primer of VHSV were designed. The pyrosequencing reaction system and conditions were optimized and the pyrosequencing method for detecting VHSV was established. [ Result] This method was only able to specifically detect the objective viruses in the eight fish viruses, and the method had the advantage of high sensitivity. The minimum detectable limit of nucleic acid was 82 copies/μL. The method was verified by detecting VHSV in 1 924 batches of samples collected from domestic and imported fishes. The detection results were consistent with that of traditional RT-PCR, and the specificity and sensitivity of the method could meet the detection requirement for aquatic animal diseases. [ Conclusion] The study provides a new detection method for monitoring and prevention and control of aquatic animal virus diseases.[Objective] The paper was to establish pyrosequencing methods for detecting viral hemorrhagic septicemia virus （VHSV）. [ Method ] One pair of PCR primers and one pyrosequencing primer of VHSV were designed. The pyrosequencing reaction system and conditions were optimized and the pyrosequencing method for detecting VHSV was established. [ Result] This method was only able to specifically detect the objective viruses in the eight fish viruses, and the method had the advantage of high sensitivity. The minimum detectable limit of nucleic acid was 82 copies/μL. The method was verified by detecting VHSV in 1 924 batches of samples collected from domestic and imported fishes. The detection results were consistent with that of traditional RT-PCR, and the specificity and sensitivity of the method could meet the detection requirement for aquatic animal diseases. [ Conclusion] The study provides a new detection method for monitoring and prevention and control of aquatic animal virus diseases.

关 键 词：Viral hemorrhagic septicemia virus （VHSV） PYROSEQUENCING Detection 

分 类 号：S941.41[农业科学—水产养殖]