李超群，张耀刚，曹得萍

(青海大学医学院)



青南藏区多房棘球绦虫系统发育学分析※

李超群，张耀刚，曹得萍※※

(青海大学医学院)

摘要目的对青南藏区流行的多房棘球绦虫基因多态性及系统发育进行初步分析。方法利用线粒体DNA细胞色素氧化酶亚单位Ⅰ(COX Ⅰ)行分子标记，以COX Ⅰ为基因marker设计引物扩增COX Ⅰ部分基因序列，将得到的11条碱基序列通过Clustal X软件进行多重序列分析比对，与13条来自GenBank的棘球绦虫COX Ⅰ序列共同构建贝叶斯进化树。结果用于分析的标本序列与来源于亚洲的中国(四川，内蒙古)、哈萨克斯坦和欧洲的法国、斯洛伐克的多房棘球绦虫型聚于一枝，位于发育树的冠部，并在这一枝内部形成梳齿状分支。结论流行于青南藏区的多房棘球蚴标本存在基因多态性，其系统发育复杂。

关键词青南藏区多房棘球绦虫细胞色素氧化酶亚单位Ⅰ系统发育

PRELIMINARY PHYLOGENETIC ANALYSIS OF

EchinococcusmultilocularIN QINGHAI SOUTHERN

PLATEAU INFERRED BY CYTOCHROME OXIDASE

SUBUNIT I GENE SEQUENCE※

Li Chaoqun，Zhang Yaogang，Wu Defang，Cao Deping※※

(Department of Joint，Qinghai University Affiliated Hospital，Xining 810001，China)

Abstract ObjectiveTo analysis the phylogeny and genetic polymorphism of Echinococcus multilocular in Qinghai southern plateau.Methods We designed and synthesized the primers of COX Ⅰ gene，and used PCR to cloned 11 sequences from hepatic sample which infected byEchinococcusmultilocular.The sequences were aligned using Clustal X，and then，Bayesian analyses were performed with 13 sequences of COX Ⅰ gene from the GenBank.Results In the phylogenetic tree，all ofEchinococcusmultilocularisolates belongs toEchinococcusmultilocularwhich originate from different places，such as China(Sichuan，Inner Mongolia )，Kazakhstan of Asia，France and Slovakia of Europe，they located on the phylogenetic tree crown，and form a comb shaped branch in this branch of internal.Conclusion There are many gene polymorphism concerningEchinococcusmultilocularisspecimens which specially popular in the southern regions of Qinghai province.

KeywordsThe south region of Qinghai provinceEchinococcusmultilocularisCytochrome oxidase subunitⅠPhylogeny

泡球蚴病(Alveolar Echinococcosis，AE)是多房棘球绦虫(Echinococcus multilocularis，Em)的继绦期幼虫寄生于人体内(多见于肝脏部位)引起的疾患。在利用DNA序列特征对虫株确定研究中，因线粒体DNA具有其母系遗传、突变率高、极少发生重组等特性，所以在分子遗传学和人类学研究上都有重要的意义[1]。本文利用COXⅠ分子标记，通过对收集的11份流行于青海省多房棘球蚴标本和GenBank下载的13条绦虫COXⅠ基因序列行比对、碱基分析，构建系统发育树，进行系统发育学研究。

1材料与方法

1.1标本采集

标本采集于青海大学附属医院和解放军第四陆军医院，共11例，类型为AE手术标本。

1.2 基因组DNA提取