Isolation of the Single Oocyst and Sequencing of ITS-1 of Eimeria intestinalis

肠艾美尔球虫单卵囊分离及ITS-1序列测定(英文)

[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.Then,according to the 18s rDNA and 5.8s rDNA sequences of Eimeria coccidia published in GenBank,a pair of specific primers was 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.

[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。根据GenBank中发表的艾美尔属球虫18s和5.8srDNA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]成功地分离出肠艾美尔球虫,其卵囊扩增出434bp清晰条带,且最低能检测出27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。