2011年济南市某污水河中肠道病毒的分离鉴定与序列分析

Identification and sequence analysis of human enteroviruses isolated from a polluted river in Jinan City,Shandong Province,2011

目的：监测济南市区地表河水中的人类肠道病毒（HEV），分析其基因特征。方法于2011年5、6、7月份对流经济南市中心城区的西泺河各采集1份河水标本，经过阴离子膜吸附的方法浓缩后，接种RD、HEp-2和L20b细胞进行病毒分离。测定分离株VP1编码区序列并进行序列分析。结果共分离到HEV 9株，其中埃可病毒6型（Echovirus 6，E6）4株，柯萨奇B2病毒（Coxsackievirus B2，CVB2）5株。VP1核苷酸序列分析显示，4株E6病毒与2010年山东省临沂市脑膜炎病例分离株的同源性较高（95．9％～96．7％），提示当地有发生E6引起的脑膜炎病例的可能性；与济南市2010年污水分离株的同源性为94．5％～99．7％。5株CVB2病毒之间有较高的核苷酸同源性（98．4％～100％），与往年山东省急性弛缓性麻痹病例分离株之间同源性为80．9％～95．2％。结论生活污水的排入河可作为HEV环境监测的采样点，我国无专门HEV病例监测系统，环境监测是了解一个地区HEV流行的重要途径。

Objective To investigate the human enteroviruses （HEVs）in Xiluo River which flows through the metro-politan area of Jinan City,and to analyze the genetic characterization of the isolates.Methods Water samples were collected monthly from the river during May to July,20 1 1 .The specimens were concentrated by anionic membrane ab-sorption and elution,and inoculated on RD,HEp-2,and L20b cell lines.The VP1 coding regions of the isolates were sequenced and phylogeneticly analyzed.Results Altogether 9 HEVs,4 echovirus 6 （E6 ）and 5 coxsackievirus B2 （CVB2）,were isolated.VP1 sequence analysis revealed the river E6 had high homology （95.9%-96.7%）with the aseptic meningitis isolate found in Linyi City in 2010,suggesting the local occurrence of E6 was associated with aseptic meningitis.River E6 also had high homology （94.5%-99.7%）with prevalent E6 strains in Jinan City in 2010.The 5 CVB2 strains were highly homologous （98.4%-100%）,which had 80.9%-95.2%homology with isolates from previ-ous acute flaccid paralysis cases in Shandong Province.Conclusion The domestic sewage-polluted river can serve as sampling site for HEV environmental surveillance,which offers an effective approach to comprehend the local HEV cir-culation in the context of no specialized surveillance system in China.