新疆部分地区屠宰场羊源细粒棘球蚴分子鉴定及其遗传多样性分析

Molecular Identification and Genetic Diversity of Echinococcus granulosus Derived from Sheep in Abattoirs in Parts of Xinjiang Uygur Autonomous Region,China

[目的]了解新疆维吾尔自治区部分规模化屠宰场羊的细粒棘球蚴感染情况及其分子遗传特征。[方法]收集屠宰场羊肝脏组织内经肉眼观察鉴定为细粒棘球蚴的包囊样本96份,全部提取基因组DNA后,基于细粒棘球绦虫cox1基因位点和nad1基因位点进行PCR扩增和测序。通过序列比对数据鉴定其基因型,构建遗传进化树解析其分子遗传特征。[结果]在cox1基因位点上,有81份样本呈PCR扩增阳性(84.37%,81/96),经序列分析鉴定,所有阳性样本均为细粒棘球蚴G1基因型(n=81),存在15个单倍型(Hap_1~15);在nad1基因位点上,有67份样本呈PCR扩增阳性69.79%(67/96),经序列分析鉴定出2种基因型,分别为细粒棘球蚴G1基因型(n=66)和G3基因型(n=1),存在19个单倍型(Hap_1~19)。种系发育分析显示,获得的细粒棘球蚴G1基因型和G3基因型序列,与国内外已报道的多种宿主源的G1基因型和G3基因型序列均处于同一个进化支。[结论]新疆部分地区羊源细粒棘球蚴呈现遗传多样性分布特征,研究结果为该地区羊包虫病的防治提供了基础数据。

[Objective]The aim of this study was to understand the infection status and molecular genetic characteristics of Echinococcus granulosus(E.granulosus)derived from sheep in some large-scale abattoirs in Xinjiang Uygur Autonomous Region,China.[Method]A total of 96 cyst samples identified as E.granulosus by macroscopic observation were collected from the liver tissues of sheep in abattoirs.Following genomic DNA extraction,PCR amplification and sequencing were performed based on the cox1 and nad1 gene loci of E.granulosus.Sequence alignment was carried out to identify the genotype.The genetic evolution tree was constructed to analyze the molecular genetic characteristics.[Result]At the cox1 gene locus,a total of 81 samples were positive in PCR amplification(84.37%,81/96).Sequence analysis identified that all positive samples were G1 genotype of E.granulosus(n=81),with 15 haplotypes(Hap_1-15)presented.At the nad1 gene locus,there were 67 PCR positive samples(69.79%,67/96).Two genotypes,G1 genotype(n=66)and G3 genotype(n=1)of E.granulosus,were identified through sequence analysis,with 19 haplotypes(Hap_1-19)presented.According to the phylogenetic analysis,the G1 and G3 genotypes of E.granulosus obtained in this study were in the same evolutionary branches as their counterparts of E.granulosus derived from the multiple host sources reported in China and abroad.[Conclusion]The E.granulosus derived from sheep in some parts of Xinjiang showed genetic diversity.Our results provide fundamental data for the prevention and control of echinocococosis in sheep in this region.