四川省骆驼源隐孢子虫的分离与虫种鉴定  

Isolation and identification of Cryptosporidium genotype and subtype from bactrian camel in Sichuan Province

作　　者：谢娜[1] 钟志军[1] 刘学涵[1] 陈维刚[2] 李威[1] 刘俊卿 李魏[1] 邓家波[2] 孙鸿雁 谢瑶 庄妤冰 徐晓阳[1] 苏怀益 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]雅安万贯公司雅安市碧峰峡野生动物园,四川雅安625007

出　　处：《中国兽医科学》2015年第4期414-418,共5页Chinese Veterinary Science

基　　金：成都大熊猫繁育研究基金会项目(CPF申2012-01);教育部"长江学者和创新团队发展计划"创新团队项目(IRT0848);2013年四川省学术带头人培养项目(83120220)

摘　　要：为调查四川省骆驼的隐孢子虫感染情况,采用蔗糖溶液漂浮法检测了成都动物园和雅安市碧峰峡野生动物园共6匹成年骆驼的新鲜粪便,通过巢式PCR扩增了阳性分离株的18SrRNA位点基因,并采用MLST方法测定了该分离株在MS1、MS2、MS3和MS16基因位点单元型;同时构建了系统发育树,鉴定了感染的隐孢子虫虫种及亚型类别。结果显示,2匹骆驼(CDBC01、SCBCO2)感染了隐孢子虫,18SrRNA序列与安氏隐孢子虫人源分离株(GenBank登录号KF271478)的相似率均为100%;经MLST分析,该2株亚型分别为A6、A5、A2、A1和A4、A4、A4、A1,与先前在骆驼与牛上的报道一致。本试验首次在四川省骆驼体内发现安氏隐孢子虫,并确定为2种隐孢子虫亚型。To examine Cryptosporidiumoocysts using the modified Sheather's sucrose flotation technique,fresh fecal specimens from six bactrian camels were collected at the Chengdu City Zoo and Ya'an City Zoo,respectively.The DNA sequence analysis of 18 SrRNA gene from the positive isolates were genotyped by the nested PCR amplification.The subtype was identified by the multilocus sequence typing(MLST)analysis.A phylogenetic tree was constructed to determine the infected species and subtypes.In result,two specimens obtained from the camels were found to be positive,and both isolates were confirmed to be Cryptosporidium andersoni with the similarities of 100% to C.andersoni reported in human(GenBank accession No.:KF271478).MLST analysis indicated that the subtypes of the C.andersoni isolates were A6,A5,A2,A1 and A4,A4,A4,A1 in the four minisatellite loci(MS1,MS2,MS3 and MS16,respectively).The above results showed that C.andersoni infecting bactrian camels in Sichuan Province was described at the first time,and two different subtypes of C.andersoni were confirmed.

关 键 词：隐孢子虫 骆驼 安氏隐孢子虫 18S RRNA MLST 种系发育分析 

分 类 号：S852.723[农业科学—基础兽医学]