一株金丝猴源毕氏肠微孢子虫的基因鉴定  

Genetic identification of a golden monkey-derived Enterocytozoon bieneusi

作　　者：李威[1] 王利勤[2] 钟志军[1] 刘学涵[1] 邓磊[1] 谢娜[1] 弓超[1] 余建秋[2] 杨俊 李奇 杨平 赵波[2] 彭广能[1] 

机构地区：[1]四川农业大学动物医学院动物疫病与人类健康四川省重点实验室,四川成都611130 [2]成都动物园成都市野生动物研究所,四川成都610081 [3]四川省雅安市名山区农业局,四川雅安625100

出　　处：《中国兽医科学》2016年第8期986-990,共5页Chinese Veterinary Science

基　　金：成都动物园(成都市野生动物研究所)科研项目(15-02)

摘　　要：采用巢式PCR法鉴定1株来自成都动物园猴源毕氏肠微孢子虫分离株(CDR01)的基因型。通过饱和蔗糖溶液漂浮法收集孢子,提取DNA后经PCR扩增ITS、MS1、MS3、MS4和MS7基因,扩增产物经测序后进行同源性和系统发育分析。结果表明,CDR01分离株的ITS、MS1、MS3、MS4和MS7基因序列与人源分离株(Gen Bank登录号:AF101200)、K-88恒河猴源分离株(Gen Bank登录号:KF305606)、秦岭羚牛源分离株1-QY25(Gen Bank登录号:KR048567)、NY-22恒河猴源分离株(Gen Bank登录号:KF305616)、人源分离株6975(Gen Bank登录号:HQ615922)的同源性均为100%。基于ITS的种系发育结果显示,CDR01与参考序列KU194595位于同一分支,ITS基因型为D型。MS1、MS3、MS4和MS7基因形成了一个MLG基因型。本研究首次报道了四川省金丝猴感染微孢子虫D型,并且用MLST法确定了该分离株的基因亚型。One Enterocytozoon bieneusi isolate CDR01 was obtained from Chengdu zoo. Spores were collected by Sheather＇s sucrose flotation technique and the genome DNA was extracted. The ITS, MSI, MS3, MS4 and MS？ genes of isolate CDROI were amplified by nested PCR and the product were sequenced followed by homology and phylogeny. In result, isolate CDROI showed lO0%homology with a homo sapiens isolate （AFIOI200）, K-88 isolate（KF305606）, I-QY25 isolate（KR048567）, NY-22 isolate（KF305616）, and 69？5 isolate （HQ615922）, respectively. The phylogenetic tree based on ITS gene showed this isolate shared the same branch with genotype D （KUl94595）.In conclusion, isolate CDROI belongs to genotype D at the ITS loci, andMSI,MS3, MS4, MSTsequencesformedaMLGtype. Thisisthefirstreport ofagoldenmonkeyinfected with genotype D of Enterocytozoon bieneusi in Sichuan province and the MLST method is used to identify the subgenotype.

关 键 词：金丝猴 毕氏肠微孢子虫 MLST 基因型 基因亚型 

分 类 号：S852.723[农业科学—基础兽医学]