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为了解山东省柯萨奇病毒B5型(Coxsackievirus B5,CV-B5)的分子进化特征,本研究对山东省2014年1~12月无菌性脑膜炎(Aseptic meningitis,AM)病例标本和环境污水监测标本进行肠道病毒(Enterovirus,EV)分离,阳性分离物采用分子生物学方法鉴定型别,并对CV-B5的VP1区序列进行系统进化树分析。结果显示,2014年AM病例中分离到69株CV-B5,占EV分离株43.95%;环境污水中分离到159株CV-B5,占非脊髓灰质炎EV分离株34.49%。上述CV-B5分离株具有季节性特征,夏秋季高峰明显。系统进化树分析表明,本研究分离株属于A、B两个基因簇,各簇之内AM和环境污水分离株之间有密切的亲缘关系,且环境污水分离株具有一定的地域聚集性。A、B基因簇的组内遗传距离分别为0.026、0.030,A基因簇与B基因簇组间遗传距离为0.085,提示CV-B5在山东地区存在2个传播链共循环,同时可能存在CV-B5跨地区传播和流行。本研究结果证明环境监测具有对EV相关疾病的预警能力。
In order to understand the molecular evolution characteristics of Coxsackievirus B5 (CV-B5) in Shandong Province, this study investigated the clinical data of aseptic meningitis (AM) from January to December 2014 in Shandong Province, Surveillance specimens were subjected to Enterovirus (EV) isolation. The positive isolates were identified by molecular biology and the VP1 region of CV-B5 was analyzed by phylogenetic tree. The results showed that 69 strains of CV-B5 were isolated from AM cases in 2014, accounting for 43.95% of EV isolates; 159 strains of CV-B5 were isolated from environmental pollutions, accounting for 34.49% of non-polio EV isolates. The above CV-B5 isolates have seasonal features with a clear peak in summer and autumn. Phylogenetic tree analysis showed that the isolates belonged to A and B clusters. There was close genetic relationship between AM and environmental isolates in each cluster, and the environmental sewage isolates had some geographical clustering. The genetic distance within the group A and B was 0.026 and 0.030, respectively. The genetic distance between the A and B gene cluster was 0.085, suggesting that CV-B5 shared two circulating loops in Shandong and there were CV- B5 spread and popularize across regions. The results of this study demonstrate that environmental monitoring has the ability to alert EV-related diseases.