诺如病毒核酸检测及病毒基因序列分析

    宋锋林 薛 芳 蔡 鹏 高玉峰 吴 刚 樊德海 张洪兵

    （辽宁出入境检验检疫局，大连 116001）

    摘要 〔目的〕通过对国境口岸输入性感染性腹泻暴发病例的快速检测、排查和病原基因序列分析，为病人的诊断、治疗和口岸突发疫情的处置提供依据，保障国境口岸的卫生安全。〔方法〕利用分子生物学技术，通过粪便样本的核酸（RNA）提取、聚合酶链反应（RT-PCR）、凝胶电泳分析以及基因序列测定、基因序列分析等过程，对造成群体腹泻的病原种类进行确认。〔结果〕9份送检的腹泻病人粪便样本中均检测到诺如病毒特异性核酸片段。经基因序列分析，确认检测到的诺如病毒基因型属基因组Ⅰ（GⅠ）。确认造成马耳他籍船舶“蓝海探索”号暴发腹泻病例的病原为诺如病毒。〔结论〕通过核酸检测等技术手段，可快速确认传染病病原种类，为疫情控制、保障国境口岸的卫生安全提供技术支持。

    关键词 诺如病毒；腹泻；核酸检测；基因序列

    〔中图分类号〕　R512.5　〔文献标识码〕　B

    Detection on the Nucleic Acid and Analysis onGene Sequencing of Norovirus   Song  Fenglin, XueFang, Cai Peng, Gao Yufeng, Wu Gang, Fan Dehai,Zhang Hongbing.Liaoning Entry-Exit Inspection and Quarantine Bureau，Dalian 116001,China.

    [Abstract]  Objective    To detect, investigate and analyze ongene sequencing of virus on imported cases of infectious diarrheaoutbreak, to provide the direct basis for patient’s  diagnose,treatment and the disposition to pestilence emergencies at thefrontier ports, and to safeguard the health and safety at thefrontier ports.  Method   The nucleicacid（RNA）extraction of the stool samples, reverse transcriptionpolymerase chain reaction（RT-PCR）, gel electrophoresis analysis,gene sequencing and the gene sequencing were conducted to confirmthe types of pathogens which caused the collective diarrhea. Result   The specific nucleic acid fragments of thenorovirus （GenomeⅠ, GⅠ）was detected in the nine stool samples, andwhich confirmed the pathogen that caused the outbreak of diarrheacases on the ship named “AZAMARA QUEST” from Malta was thenorovirus（GenomeⅠ, GⅠ）.   Conclusion  Nucleic aciddetection techniques can quickly identify types of infection forepidemic control, health and safety protection at the frontierports.

    [Key words]   Norovirus; Diarrhea;Nucleic acid detection; Sequence analysis