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当前位置: 首页 > 文章 > 病毒性出血性败血症病毒焦磷酸测序检测技术的建立 中国预防兽医学报 2017 (7) 564-568
Position: Home > Articles > Development of pyrosequencing methods for detecting viral hemorrhagic septicemia virus Chinese Journal of Preventive Veterinary Medicine 2017 (7) 564-568

病毒性出血性败血症病毒焦磷酸测序检测技术的建立

作  者：
尹伟力;刘瑶;黄微;张四化;岳志芹;孙涛
单  位：
山东出入境检验检疫局技术中心;烟台出入境检验检疫局;荣成出入境检验检疫局;武汉市农业检测中心;武汉市动物疫病防控中心
关键词：
病毒性出血性败血症病毒;焦磷酸测序;检测
摘  要：
为建立病毒性出血性败血症病毒(VHSV)焦磷酸测序检测技术,本研究通过对VHSV靶基因特异性序列的同源性分析,设计出适合焦磷酸测序特异性扩增和测序的引物,对焦磷酸测序反应体系和反应程序进行优化,建立了VHSV焦磷酸测序检测方法。结果表明,该方法仅对目的病毒基因扩增,而对其它病毒检测为阴性,表明该方法特异性好;该方法最低检出核酸量为82拷贝/μL,灵敏度高。选取国内采集与进口的鱼类样本共计1924批次进行VHSV检测,结果显示,焦磷酸测序检测方法检测结果与常规RT-PCR一致,该方法的特异性和灵敏度可以满足水生动物疫病检测的需要。
译  名：
Development of pyrosequencing methods for detecting viral hemorrhagic septicemia virus
作  者：
YIN Wei-li;LIU Yao;HUANG Wei;ZHANG Si-hua;YUE Zhi-qin;SUN Tao;Yantai Entry-exit Inspection and Quarantine Bureau;Rongcheng Entry-exit Inspection and Quarantine Bureau;Wuhan Testing Center for Agricultural;Wuhan Center for Animal Disease Control and Prevention;Technical Center of Shandong Entry-Exit Inspection and Quarantine Bureau;
关键词：
viral hemorrhagic septicemia of salmonids virus(VHSV);;pyrosequencing;;detection
摘  要：
To develop pyrosequencing methods for detecting viral hemorrhagic septicemia virus, one pair of PCR primers and one pyrosequencing sequencing primer of VHSV were designed, then the pyrosequencing reaction system and conditions were optimized and the pyrosequencing method for detection of VHSV was established. 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 and the minimum detection limit of nucleic acid is 82 copes/μL. The method was verified by the detection on VHSV in 1924 batches of the samples collected from domestic and imported fishes. The detection results were in consistence with that of traditional RT-PCR, and the specificity and sensitivity of the method could meet the requirement for detection of aquatic animal diseases.

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