环境污水中诺如、轮状、星状病毒的检测和分子流行病学研究

Detection and Molecular Epidemiology of Norovirus,Rotavirus and Human Astrovirus in Domestic Sewage

为分析城市污水中诺如病毒（Norovirus,NoV）、轮状病毒（Rotavirus,RV）、人类星状病毒（Human Astrovirus,HAstV）的分子流行病学特征,探索环境监测技术在病毒性胃肠炎疾病防控中的作用,本研究在山东省3个城市建立环境污水监测点,从7份分别采自2009～2015年的污水标本中提取RNA,通过逆转录-聚合酶链反应扩增其中的NoV、RV、HAstV的核酸片段,扩增产物通过TA克隆后转化大肠杆菌JM109并进行序列测定,根据获得的序列分析其型别构成和系统发生特征。共检测出特异性序列210条,分属于6个NoV I基因型、4个NoV II、3个RV G基因型、3个RV P基因型和4个HAstV血清型。GI.2、GII.4、G9、P[8]和HAstV-1是各病毒检测中最常见的基因型。系统发生分析显示GI.3、GI.6、GII.4、G9、P[8]、HAstV-1和HAstV-4基因型内又分为多个传播链。研究结果表明生活污水中含有多种多样的胃肠炎病毒信息,环境监测是监测病毒区域性流行的重要途径之一。

To assess the significance of environmental surveillance in the control and prevention of viral gastroenteritis,we analyzed the molecular characteristics of norovirus（NoV）,rotavirus（RV）,and human astrovirus（HAstV）,sequences detected in domestic sewage.Environmental sewage monitoring sites were set up in three cities in Shandong,China.RNA was extracted from seven sewage samples collected each year from 2009 to 2015.RT-PCR detection of norovirus,rotavirus,and human astrovirus was performed.Positive PCR segments were cloned into a T-vector,transformed and sequenced,and genotyping and phylogenic analysis performed.A total of 210 viral sequences belonging to 6NoV I,4NoV II,3RV G,3RV P and 4HAstV were obtained.GI.2,GII.4,G9,P[8],and HAstV-1were the most frequently detected types.Phylogenetic analysis revealed multiple transmission chains in the genotypes of GI.3,GI.6,GII.4,G9,P[8],HAstV-1,and HAstV-4.The results showed not only that sewage contains dramatic information regarding gastroenteritis viruses,but also that environmental surveillance is an important approach in monitoring the regional circulation of specific viruses.