[目的]建立一种有效的鉴定球虫种类分子生物学方法.[方法]采用单卵囊分离技术,分离肠艾美尔球虫,提取卵囊基因组DNA.根据GenBank中发表的艾美尔属球虫18S rDNA和5.8S rDNA序列,设计特异性引物,扩增内转录间隔区1(ITS-1),PCR产物直接测序.[结果]成功分离出肠艾美尔球虫,PCR扩增出434 bp清晰条带,且最低能检测出27个孢子化卵囊 .[结论]该研究结果为球虫虫种及虫株的准确鉴定奠定了基础.%[ Objective ] The aim was to establish a molecular biological method for identifying coccidium species. [ Method ] First, the pure species of Eimeria intestinalis was isolated by using single-oocyst isolation technique, and oocyst genome DNA was extracted. Then, according to the l8s rDNA and 5.8s rDNA sequences of Eimeria Coccidia published in GenBank, a pair of specific primers were designed and synthesized to amplify the internal transcribed spacer 1 ( ITS-1 ). Finally, the PCR products were sent for sequencing. [ Result ]. The pure species of E. intestinalis was isolated and the result of agarose gel electrophoresis showed that the PCR product was 434 bp, and at least 27-sporulated oocysts could be detected. [ Conclusion ] The research will provide a basis for accurate identification of coccidium species and strains.
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