畜牧兽医学报2012，Vol.43Issue(6)：1003-1008,6.
斯氏艾美耳球虫ITS1-5.8S rRNA-ITS2序列的克隆及PCR检测方法的建立
Cloning and Analysis of ITS1-5.8S rRNA-ITS2 of Eimeria stiedai and Development of PCR Diagnostic Assay Based on the Fragment
摘要
Abstract
18S rRNA and 28S rRNA sequences of several Eimeria species from chickens and squirrels were aligned to design common primers for Eimeria parasites from various hosts based on the conserved sequences of both 3' end of 18S rRNA and 5' end of 28S rRNA. 1 178 bp of the ITS1-5. 8S rRNA-ITS2 complete sequence of E. stiedai, including 423 bp of ITS1, 155 bp of 5. 8S rRNA, and 600 bp of ITS2, was firstly cloned with the common primers and genomic DNA of oocysts of LY isolate as templates. In contrast to 5. 8S rRNA fragment, ITS1 and ITS2 sequences of E. stiedai LY isolate was more variable, and less than 60% of ITS1 and ITS2 sequences of LY isolate was identical to those of Eimeria species in chickens and other rodent hosts. A sensitive and specific PCR diagnostic assay based on the ITS1-5. 8S rRNA-ITS2 sequence was developed to identify E. stiedai, one of high pathogenic species from rabbits by designing specific primers for E. stiedai at the mutative sites of ITS1 and ITS2. These findings will provide a powerful tool for clinical differentiation of high pathogenic Eimeria species in rabbits and revealing population genetic characteristics of rabbit coccidia.关键词
兔/斯氏艾美耳球虫/ITS1-5.8S rRNA-ITS2序列/PCR检测Key words
rabbits/ Eimeria stiedai/ ITS1-5. 8S rRNA-ITS2 sequence/ PCR diagnostic assay分类
农业科技引用本文复制引用
闫文朝,韩利方,张龙现,索勋,薛帮群,王帅..斯氏艾美耳球虫ITS1-5.8S rRNA-ITS2序列的克隆及PCR检测方法的建立[J].畜牧兽医学报,2012,43(6):1003-1008,6.基金项目
国家自然科学基金项目(31001058) （31001058）
国家兔产业技术体系项目(nycytx-44)资助 （nycytx-44）
