山羊源枝睾阔盘吸虫的形态和分子鉴定

The morphological and molecular identification of Eurytrema cladorchis isolated from goats

为鉴定采自西昌市6只山羊体内的小型吸虫的种类,本研究采用传统的形态学观察和PCR方法对其进行种类鉴定。经形态学观察,发现采自6只山羊体内的吸虫与枝睾阔盘吸虫形态相似;对6个样本虫株cox1基因部分序列进行PCR扩增和序列测定,并用cox1序列构建其与其他吸虫的系统发育关系。测序结果显示所获得的吸虫cox1基因片段大小为429 bp,6个虫株的同源性为97.7%~99.8%;构建的NJ系统发育树显示6个虫株与胰阔盘吸虫的亲缘关系最近。因此,结合形态学观察结果,将分离的6个小型吸虫虫株均鉴定为枝睾阔盘吸虫。研究结果为枝睾阔盘吸虫病的分子诊断、分子流行病学调查的进一步研究奠定了基础。

In order to identification the species of flukes isolated from six goats in Xichang city, morphological observation and PCR method were used in this study. All the flukes isolated from goats were identified as Eurytrema cladorchis by the morphological observation; the partial sequences in cox1 gene of six fluke samples were amplified by PCR and sequenced, and the phylogenetic tree based on cox1 was constructed using MEGA 5.0 software. The results showed that the length of partial sequence in cox1 gene was 429 bp, and the homology among these sequences was 97.7%-99.8%. The phylogenetic tree constructed by NJ method based on cox1 gene showed that the six samples had a closest relationship with the Eurytrema pancreatic. Therefore, the six fluke isolates were identified as E.cladorchis by morphological and molecular studies. The results provided the foundation for further research on molecular classification, epidemiological survey of Eurytrema cladorchis.