Objective: To understand the genotypes and nucleotide polymorphisms of Echinococcus granulosus metacestode from humans and sheep in Tianjun region, Qinghai Province.
Methods: The specific primers were designed according to the cox1 and nad1 genes of E. granulosus mitochondrial genome sequences accessed by GenBank. The primers were used to detect the cyst samples from 16 sheep and 2 humans infected with E. granulosus in Tianjun region of Qinghai Province by PCR, then the PCR amplification products were sequenced, the genotypes and nucleotide polymorphisms of the cox1 and nad1 genes were analyzed.
Results: The 18 isolated samples all belonged to E. granulosus G1 genotype. Among all the isolates, 9 haplotypes existed in the cox1 gene with 16 nucleotide mutation sites, and there were 0 to 5 nucleotide differences with the highest variation rate of 0.31%, whereas 7 haplotypes occurred with 15 nucleotide mutation sites, and there were 1 to 8 nucleotide differences with the highest variation rate of 0.89% for the nad1 gene.
Conclusions: The epidemic genotype of E. granulosus is G1 in humans and sheep in Tianjun region of Qinghai Province, and the nucleotide polymorphisms of the cox1 gene were more abundant than those of the nad1 gene, and the resolution of the nucleotide polymorphisms of cox1 gene is higher than that of the nad1 gene used in E. granulosus isolates.
[摘要] 目的 解析青海天峻地区人和绵羊细粒棘球蚴流行株的基因型及基因多态性。方法 根据GenBank公布的细 粒棘球绦虫线粒体基因组序列, 分别针对cox1和nad1基因设计特异性引物, 对16份绵羊源和2份人源细粒棘球蚴分离 株样品进行PCR扩增、测序及cox1和nad1基因核苷酸序列多态性和单倍型分析。结果 18株细粒棘球蚴分离株均属于 G1型。其中cox1基因单倍型共有9种, 变异位点16个, 单倍型序列之间碱基差异0~5个, 变异率达0.31%; nad1基因单倍 型共有7种, 涉及变异位点共15个, 单倍型序列之间碱基差异1~8个, 变异率达0.89%。结论 青海天峻地区人和绵羊细 粒棘球蚴流行株为G1型, 其cox1基因多态性较nad1基因多态性丰富; cox1基因用于细粒棘球蚴分离株间核苷酸多态性 分析的分辨率较高。.
Keywords: Echinococcus granulosus; Genotyping; Nucleotide polymorphism; cox1 gene; nad1 gene.