新疆塔城地区马梨形虫感染的分子生物学诊断及分析

Diagnosis and Analysis of Molecular Biology of Horse Equine Piroplasma Infection in Tacheng,Xinjiang

采用常规血涂片镜检和PCR技术对新疆塔城地区疑似梨形虫感染的病马进行检测。通过18S rRNA基因通用引物扩增出目的基因,克隆测序后,与GenBank公布的梨形虫18S rDNA参考序列对比分析同源性,并构建系统进化树,确定梨形虫的种属。血涂片鉴定结果显示,红细胞中发现圆环形虫体,具有马梨形虫的主要特征。分子生物学鉴定发现,18S rDNA序列与马泰勒虫美国分离株核苷酸序列同源性为98.3%;遗传进化分析发现,该虫株与已知的泰勒属在同一大枝上,与马泰勒虫美国分离株、马泰勒虫西班牙分离株有较近的亲缘关系。

The routine blood smear and PCR technique were used to detect the disease of horses in Tacheng area of Xinjiang.Target gene was amplified by PCR using 18 S rRNA gene universal primers.After cloning and sequencing,the identity was analyzed between the sequenced and 18 S rDNA reference sequence downloaded from GenBank and phylogenetic tree was constructed among piroplasmaspecies.Blood smear tests show that the annular shaped worm with the main characteristics of equine piroplasmid were found in red blood cells.The identification of molecular biology based on 18 S rDNA sequences showed that the isolate shared 98.3%identities in nucleotide sequence with Theileria equi strain.Genetic evolution analysis found that the isolate has closer genetic relationship with US Theileria equi strains,Theileria equi Spanish isolates,which was located in the same Branch in phylogenetic tree.