SSR-PCR和常规PCR检测不同地区并殖吸虫遗传变异的比较研究

目的以SSR-PCR和常规PCR对浙江省和福建省5个不同地区并殖吸虫进行遗传变异检测,比较两者对并殖吸虫的基因水平分类的差异。方法采用微卫星锚定PCR（SSR-PCR）对基因组DNA进行PCR扩增,扩增产物按Nei的方法计算遗传距离,并应用SPSS软件构建系统进化树;采用常规PCR扩增线粒体细胞色素C氧化酶亚单位（COI）及核糖体DNA第二间区（ITS2）的基因序列,测序后用BoiEdit软件分析同源性,用MEGA软件构建种系发生树。结果不同地区并殖吸虫标本的SSR-PCR产物呈多态性,浙江金华、浙江永嘉、福建周宁与福建平和、福建政和的标本存在明显的差异。前两者的遗传距离最近,为0.3333,浙江金华与福建平和的标本遗传距离最远,为0.8667。常规PCR基因序列分析显示浙江金华、浙江永嘉、福建周宁的标本之间在种系发生树上比较接近,浙江金华和浙江永嘉的标本最为接近。结论利用SSR-PCR和常规PCR进行并殖吸虫的遗传变异分析结果基本一致。SSR-PCR是一种比常规PCR更方便的并殖吸虫遗传变异研究方法。

A comparison between SSR-PCR and normal PCR technology in detection of Paragonimus in different areas

The aim was to detect the genetic variation of Paragonimus in 5 areas in Zhejiang Province and Fujian Province with technologies of SSR-PCR and normal PCR,so as to compare the differences on the gene horizontal classification.Our method was to amplify the genome DNA with the technology of simple sequence repeat-anchored PCR（SSR-PCR）,and count the genetic distance with the method of Nei,and construct the dendrogram with SPSS software.This study was also corduct to amplify COI gene and ITS2 gene by normal PCR,analyze homology through BoiEdit,and construct stammbaum through MEGA.The SSR-PCR product of the Paragonimus speciman in different areas was polymorphic.There were obvious differences between Jinhua in Zhejiang Province,Yongjia in Zhejiang Province,Zhouning in Fujian Province and pinghe and zhenghe in Fujian province.The genetic distance of the first two is the nearest（0.3333）,and the specimen in Jinhua is the farthest to the Pinghe,with each other（0.8667）.According to the normal PCR gene sequencing,specimen in Jinhua,Yongjia,and Zhouning are near in the phylogenetic tree,and the specimen in Jinhua is nearest to that in Yongjia.The results are similar on genetic variation of Paragonimus with technologies of SSR-PCR and normal PCR.It＇s indicated that SSR-PCR was a more convenient method on studying the genetic variation of Paragonimus.