应用ISSR-PCR技术对我国11株并殖吸虫遗传变异的研究

目的对我国3省9地11株并殖吸虫进行遗传变异研究,为并殖吸虫的基因分类提供依据方法采用简单重复序列间PCR(ISSR-PCR)技术对11株并殖吸虫基因组DNA进行扩增,根据扩增产物电泳结果计算遗传距离并绘制系统进化树。结果江西武宁、浙江兰亭、湖北温泉和湖北赤壁等4株与其余7株并殖吸虫扩增产物之间存在较明显的差异;湖北温泉和湖北赤壁株遗传距离为0.181 8;湖北五峰(a)、湖北五峰(b)和湖北走马(b)株之间扩增产物遗传距离在0.125~0.18之间;湖北走马(a)和湖北五里株间遗传距离为0.153 9;湖北红坪和湖北十堰株遗传距离为0.156 6。结论11株并殖吸虫在基因水平分为两大类:1)武宁株、兰亭株、赤壁株和温泉株,属卫氏并殖吸虫;2)红坪株、走马(a)株、走马(b)株、五峰(a)株、五峰(b)株、五里株和十堰株,属斯氏狸殖吸虫。湖北省存在卫氏并殖吸虫流行区和斯氏狸殖吸虫流行区。采用并殖吸虫不同虫期(成虫和囊蚴)标本及其不同保存方法对ISSR-PCR结果无影响。

Genetic variation of 11 strains of Paragonimus detected by the inter-simple sequence repeat PCR (ISSR-PCR) technique

Objective To study the genetic variation of 11 strains of Paragonimus from 9 districts of 3 provinces in China and provide genetic evidence for the classification of Paragonimus.Methods Paragonimus isolates were obtained from seven areas in China.Total genomic DNA was used as template and amplified by inter-simple sequence repeat PCR(ISSR-PCR) technique.The genetic distances among these isolates were measured according to the amplified products and the corresponding dendrogram was constructed.Results The distance between the 4 isolates of Hubei-Wenquan,Jiangxi-Wuning,Zhejiang-Lanting and Hubei-Chibi and the other 7 isolates(Hubei-Shiyan,Hubei-Wufeng,Hubei-Wuli,Hubei-Zouma,Hubei-Hongping) was significant;the genetic distance between Hubei-Wenquan isolate and Hubei-Chibi iaolate was 0.181 8.The distances among Hubei-Wufeng(a) isolate,Hubei-Wufeng(b) isolate and Hubei-Zouma(b) isolate ranged from 0.125 to 0.18.The distances between Hubei-Zouma(a) and Hubei-Wuli,and Hubei-Hongping and Hubei-Shiyan were 0.153 9 and 0.156 6 respectively.Conclusion 11 strains of Paragonimus involved in this article can be divided into two groups based on genetic level:1) the isolates from Jiangxi-Wuning,Zhejiang-Lanting,Hubei-Wenquan and Hubei-Chibi,were in the group of Paragonimus westermani;2) the isolates from Hubei-Wufeng(a),Hubei-Wufeng(b),Hubei-Zouma(b),Hubei-Zouma(a),Hubei-Wuli,Hubei-Hongping and Hubei-Shiyan,were in the group of Pagumogonimus skrjabini.There were two species of Paragonimus westermani and P.skrjabini in Hubei Province.Study with different stages of samples and different preservative methods of Paragonimus showed no effect on the amplification result.