摘要
目的对一起南京高校诺如病毒急性胃肠炎疫情进行病原基因测序分析,以确认其病原体,并对其基因进行分型研究。方法用荧光定量PCR法对采集的50份疫情病例样本进行诺如病毒RNA检测,对阳性标本采用诺如病毒的开放读码框架2（ORF2）基因N/S区域（N-terminus/shell region）进行RT-PCR扩增、测序,结合诺如病毒Ⅱ型各基因型参考株序列进行序列比对和进化树分析。结果荧光定量PCR检测的50份疫情样本中,诺如病毒Ⅱ型核酸阳性12份,其中5份病毒用RT-PCR扩增出条带,其N/S区域序列与2012年全球流行的新型诺如病毒GⅡ.4/Sydney变异株同源性高达97.7%~98.8%。结论序列测定结果证实这是南京首次由GⅡ.4/Sydney诺如病毒变异株引起的暴发疫情,在日常监测样中也发现该型病毒散发感染,提示该型病毒将是本市今后防控监测的重点。
Objective To sequence and analyze the pathogen gene in one case of campus acute gastroenteritis outbreak in Nanjing by reverse transcriptase- polymerase chain reaction（ RT- PCR） assay,in order to determine the pathogen,and study subgenotypes. Methods Real- time reverse transcription- PCR was carried out on RNA isolated from the 50 samples of the gastroenteritis outbreak. The positive samples were then amplified by reverse transcription- polymerase chain reaction（ RT- PCR）by using primers designed based on the N / S region（ N- terminus / shell region）. The PCR production were sequenced and subsequently submitted to phylogenetic analysis. Results Twelve samples were found GⅡ virus positive by real- time RT- PCR analysis. Homology analysis revealed that the sequences of 5 specimens were homogeneous with genogroup GⅡ. The N / S region of 5 samples amplified in seminested RT- PCR shared a 97. 7% ~ 98. 8% similarity in nucleotide sequences with GⅡ. 4 / Sydney variant. Conclusion The results obtained in this study demonstrates that the etiological agent in the outbreaks of acute gastroenteritis in Nanjing was GⅡ. 4 / Sydney variant,and we will focus on the prevention and control monitoring in Nanjing.
出处
《中国卫生检验杂志》
北大核心
2014年第24期3501-3504,共4页
Chinese Journal of Health Laboratory Technology
基金
青奥会食品安全保障关键技术应用与示范项目(2011-BAK21B00)
南京市卫生青年人才培养工程项目(QR-X11039)
关键词
诺如病毒
遗传组
基因型
序列分析
Norovirus
Genogroup
Genotype
Sequential analysis
