应用RAPD技术对我国11个地域株并殖吸虫遗传变异的初步探讨

目的对来自中国3省9地11个地域株并殖吸虫进行遗传变异研究,明确它们的亲缘关系,为并殖吸虫的分类提供依据,尤其为湖北省并殖吸虫流行区划分提供分子生物学依据;探讨采用不同虫期标本及其不同保存方法对该研究的影响。方法采用随机扩增多态性DNA(RAPD)技术,对11个地域株并殖吸虫进行PCR扩增,根据扩增产物琼脂糖凝胶电泳的带型,计算各株并殖吸虫的遗传距离,并绘制系统进化树。结果RAPD技术将11个地域株并殖吸虫分为3类:①江西武宁、浙江兰亭、湖北温泉和湖北赤壁;②湖北走马(a)、湖北走马(b)、湖北五峰(a)、湖北五峰(b)、湖北十堰和湖北五里;③湖北神农架。结论11个地域株并殖吸虫在基因水平上存在差异,其中有种间和种内差异。我国湖北、江西和浙江3省存在3种并殖吸虫:卫氏并殖吸虫、斯氏并殖吸虫和一种尚未定种的神农架并殖吸虫;湖北存在除卫氏、斯氏并殖吸虫外不明种的神农架并殖吸虫。采用不同虫期(成虫和囊蚴)标本及其不同保存方法不影响遗传变异研究的结果。

Preliminary analysis of genetic variation of 11 populations of Paragonimus in China by random amplified polymorphic DNA technique

To analyze the genetic variation of 11 populations of Paragonimus collected from 9 districts of 3 provinces in China and to investigate the phylogenetic relationship among them, in order to offer biological evidence for Paragonimus prevalense in Hubei province,the genomic DNAs of 11 strains were amplified by random amplified polymorphic DNA (RAPD) techngiue and the genetic distances of the Paragonimus were calculated according to the types of the amplified products on the agarose gel electrophoresis. Then, the phylogenetic tree was thereby mapped. The populations of Paragonimus observed in the present study could be divided into 3 groups: (1) isolates from Jianxi-Wuling, Zhejiang-Lanting, Hubei-Wenquan and Hubei-Chibi, (2) isolates from Hubei-Zouma(a), Hubei-Zouma(b), Hubei-Wufeng(a), Hubei-Wufeng(b), Hubei-Shiyan and Hubei-Wuli, (3) isolates from Hubei-Shennongjia. It is concluded that 11 populations of Paragonimus from different areas are genetic differences and there are inter-species and intra-species differences among them. Three species of Paragonimus: P. westermani, P. skrjabini and an uncertain species named P. sp. SNJ exist in Hubei, Jiangxi and Zhejiang province of China. While an uncertain species named P. sp. SNJ lives in Hubei province besides of P. westermani and P. skrjabini. There are no genetic variations in the samples from different stages and different preservative methods of Paragonimus.