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目的 研究中国日本血吸虫各地域株基因差异。 方法 收集湖南、安徽、湖北、四川及江苏 5省的阳性钉螺 ,将逸出的日本血吸虫尾蚴感染昆明小鼠 ,42d后 ,用灌注法获取成虫 ,用PCR扩增成虫线粒体NADH (还原型烟酰胺腺嘌呤二核苷酸 )脱氢酶 1(ND1)和细胞色素C氧化酶 1(CO1)基因片段 ,克隆于质粒载体后测序并用MEGA软件分析。 结果 各地域株日本血吸虫成虫的ND1和CO1基因长度分别为 476bp和 10 33bp ,ND1和CO1基因序列分别存在 2个单倍体 (Ⅰ型 ,Ⅱ型 ) ,其差异分别为 4.0 %和 3.4%。从分子发生树看 ,各地域株日本血吸虫ND1和CO1基因分别存在 2个不同基因型。在ND1基因中呈现Ⅰ型的 ,CO1基因中也为Ⅰ型 ,反之 ,均为Ⅱ型。 结论 研究区域湖南、安徽、湖北、四川及江苏 5省的日本血吸虫株ND1和CO1基因存在 2个不同基因型。
Objective To study the genetic differences among different regions of Schistosoma japonicum in China. Methods Positive snails were collected from five provinces of Hunan, Anhui, Hubei, Sichuan and Jiangsu Provinces. Kunming mice were infected with the secreted Schistosoma japonicum cercariae. After 42 days, adult worms were obtained by perfusion. The mitochondria NADH (reduced nicotinamide) Adenine dinucleotide dehydrogenase 1 (ND1) and cytochrome C oxidase 1 (CO1) gene fragments were cloned in a plasmid vector and sequenced and analyzed using MEGA software. Results The length of ND1 and CO1 genes of adult Schistosoma japonicum were 476 bp and 10 33 bp, respectively. There were two haplotypes (type Ⅰ and type Ⅱ) in ND1 and CO1 gene sequences, with the difference of 4.0% and 3.4% respectively. According to the molecular phylogenetic tree, there are two different genotypes of Schistosoma japonicum ND1 and CO1 gene in each region. In the ND1 gene showed type Ⅰ, CO1 gene is also type Ⅰ, on the contrary, are type Ⅱ. Conclusion There are two different genotypes of ND1 and CO1 genes in Schistosoma japonicum strains in Hunan, Anhui, Hubei, Sichuan and Jiangsu provinces.