长春地区绵羊和兔隐孢子虫分子流行病学调查  

Cryptosporidiosis Molecular Epidemiology of Sheep and Rabbits in Changchun Area

作　　者：门静涛[1] 张西臣[1] 于师宇 李建华[1] 宫鹏涛[1] Xiao Lihua 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]福清出入境检验检疫局,福清350300 [3]Division of Parasitic Diseases, Centers for Disease Control and Prevention, U. S. Department of Health and HumanServices, Atlanta 30341, U. S. A

出　　处：《吉林农业大学学报》2009年第4期447-451,共5页Journal of Jilin Agricultural University

基　　金：国家自然科学基金项目[(2002)30210403203];国家高技术研究发展(863)计划项目(2006AA06Z414);福建出入境检验检疫局项目(FK2006-02)

摘　　要：采用抗酸染色法分别对长春地区287份兔粪便样品和167份绵羊粪便样品进行隐孢子虫检查,确认阳性的样品用巢式PCR-RFLP方法扩增隐孢子虫Small-Subunit rRNA基因,选择限制性内切酶SspⅠ和VspⅠ分别对该目的片段单酶切,PCR产物测序后进行RFLP及系统发育分析。结果共检出97份兔粪便和45份羊粪便含有隐孢子虫卵囊,感染率为33.8%和26.9%。巢式PCR-RFLP检测发现由绵羊和兔粪便中分离的隐孢子虫分别是C.parvum和C.bovis,系统发育分析显示隐孢子虫虫种形成了2个群,一个群包括C.parvum,C.bovis,C.baileyi,C.meleagridis,C.felis和C.wrairi,另一个群包括C.andersoni,C.muris和C.serpentis。Two hundreds and eighty seven rabbit feces and 167 sheep feces were collected and detected by Acid-fast staining. The positive samples were analyzed by nested PCR- RFLP, in which a approximately 830 bp fragment was amplified in positive samples, and the fragment was digested by Ssp I and Vsp I , with the help of RFLP and DNA sequencing analysis. Cryptosporidium separated from sheep and rabbits in Changchun area was identified at molecular level. The results showed that the 97 rabbit feces and 45 sheep feces were positive by Acid-fast staining and the infected ratios were 33.8 % and 26.9% respectively. The nested PCR - RFLP result indicated that two species Cryptosporidium or C. parvum （separat- ed from rabbits）and C. bovis （separated from sheep） were found. Phylogenetic analysis showed that the Cryptosporidium species formed two clades. One clade contained C. parvum, C. baileyi, C. bovis C. meleagridis, C. f ells and C. wrairi. The other clade contained C. andersoni, C. muris and C. serpentis

关 键 词：隐孢子虫 分子流行病学 巢式PCR RFLP 

分 类 号：S852.72[农业科学—基础兽医学]