猴源贾第虫新亚型B11的基因鉴定  

Genetic identification of new subtype B11 of Giardiafrom monkey

作　　者：李威[1] 钟志军[1] 屈羽[2] 刘学涵[1] 谢娜[1] 邓家波[2] 周紫峣[1] 刘俊卿 程云 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,四川雅安625014 [2]成都动物园,四川成都610081 [3]四川雅安万贯公司碧峰峡野生动物园,四川雅安625007

出　　处：《中国兽医科学》2015年第2期156-160,共5页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31272620);成都动物园项目(2014-04)

摘　　要：运用饱和蔗糖溶液漂浮法收集了26份雅安市碧峰峡野生动物园不同野生动物的粪便寄生虫卵囊,提取DNA,经巢式PCR扩增β-giardin和tpi基因,扩增产物经测序后进行同源性和系统发育分析。结果显示,环尾狐猴(YARTL01)和猕猴(YARM01)感染贾第虫,其中YARTL01分离株为一个新基因亚型,被命名为B11;YARM01分离株为蓝氏贾第虫B1亚型。YARTL01分离株的β-giardin基因序列与环尾狐猴分离株6LC(GenBank登录号HQ616629)的相似率为99.4%,tpi基因序列与猕猴分离株34538(GenBank登录号JX000563)的相似率为98.6%。种系发育分析显示,YARTL01在β-giardin基因位点与聚集体B处于同一分支,而在tpi基因位点,此分离株单独成一分支。YARM01的β-giardin基因序列和tpi基因序列分别与河狸分离株Be1(GenBank登录号EU014382)、猕猴分离株36395(GenBank登录号KC441076)的相似率达到100%;在进化树中,YARM01分离株在β-giardin基因位点与聚集体B处于同一分支,在tpi基因位点此分离株与聚集体B1处于同一个进化分支。本研究是首次报道四川省猴感染贾第虫,其中YARTL01分离株为一个新基因亚型。Fresh fecal samples from 26 different wild animals were obtained at the Bifengxia Zoo of Ya'an of Sichuan,China.Parasite oocysts of each fece were collected by the Sheather's sucrose flotation technique,and their genomic DNAs were extracted.Theβ-giardin and tpi genes were amplified fromGiardiaby PCR.Phylogenetic analysis was used to identify the species and subtypes.In result,only a ring-tailed lemur(YARTL01)and a rhesus macaque(YARM01)were diagnosed positive.The maximum similarities of nucleotide homology in theβ-giardin and tpi loci were 99.4%and 98.6%,compared to those of 6LC isolate of YARTL01 and 34538isolate of YARTL01,respectively.The maximum similarity of nucleotide homology in both loci in YARM01 were 100%,compared to Be1 and 36395isolates.Phylogenetic analysis indicated that two positive isolates shared the same evolution branch with assemblage B in theβ-giardin gene,while in the tpi gene,the isolate of YARM01 yielded the same evolution branch to subtype B1,and the isolate of YARTL01 grouped a cluster separately and was named subtype B11.In conclusion,this is the first report on Giardiainfection in monkey in Sichuan Province and a new subtype B11 is identified.

关 键 词：贾第虫 巢式PCR β-giardin基因 tpi基因 种系发育分析 亚型 猴 

分 类 号：S852.7[农业科学—基础兽医学]