18S rRNA基因巢式PCR-RFLP鉴定吉林、大庆地区断奶前奶牛隐孢子虫分离株  

Chracterization of Cryptosporidium spp from preweaned calves of Jilin and Daqing area by 18S rRNA gene nested PCR-RFLP

作　　者：苏艳[1] 白光彦 孙喜东[3] 刘妍[4] 王春仁[5] 张静[1] 宋军澎[1] 尹继刚[1] 

机构地区：[1]吉林大学人兽共患病研究所,人兽共患教育部重点实验室,吉林长春130062 [2]吉林市丰满区小白山乡畜禽防疫服务中心,吉林吉林132108 [3]解放军第208医院,吉林长春130062 [4]天津工业大学外国语学院,天津300387 [5]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《中国兽医学报》2011年第3期347-351,共5页Chinese Journal of Veterinary Science

基　　金：国家“十一五”传染病重大专项基金资助项目(2008ZX10004-011)

摘　　要：利用18S rRNA巢式聚合酶链反应(Nested PCR)-限制片段长度多态性(Restriction fragment length polymorphism,RFLP)鉴定吉林、大庆地区牛源隐孢子虫分离株。采取吉林、大庆地区断奶前犊牛粪便,提取DNA后经18S rRNA基因巢式PCR扩增,扩增产物测序后用Blast和MEGA4.0软件进行同源性和系统发育树分析。同时扩增产物分别用SspⅠ、VspⅠ和MboⅡ酶切后进行RFLP分析。通过18S rRNA基因PCR-RFLP分析和测序比对分析表明,吉林分离株包括2种隐孢子虫,分别为C.bovis和C.ryanae。大庆分离株包括3种,分别为C.bovis、C.ryanae和C.andersoni。Cryptosporidium SPP isolated from feces of preweaned calves in Jilin and Daqing area were identified by 18S rRNA gene nested PCR-RFLP.The genomic DNA of Cryptosporidium was extracted from fecal samples and amplified by using the 18S rRNA gene nested PCR-RFLP assay,and Blast and MEGA4.0 softwares were used to analyze their homology and phylogeny.Meanwhile,their amplified products were digested with restriction enzymes SspⅠ,VspⅠand MboⅡ,respectively.All the digested products were analyzed with RFLP assay.As demonstrated by 18S rRNA gene analysis,the Jilin isolates included C.bovis,and C.ryanae.While the Daqing isolates included C.bovis, C.ryanae and C.andersoni.

关 键 词：安氏隐孢子虫 牛型隐孢子虫 C.ryanae 18S RRNA 巢氏PCR-RFLP 

分 类 号：S852.723[农业科学—基础兽医学]