云南6个地域带绦虫随机扩增DNA多态性分子鉴定分析研究

Analysis by morphology and PCR on Taenia of RAPD from 6 regions of Yunnan province

目的分别对云南省怒江、迪庆、大理、临沧、楚雄和西双版纳的14株带绦虫标本进行分子鉴定分析。方法在形态学鉴定的基础上取13株牛带绦虫标本株、1株猪带绦虫株、2株亚洲带绦虫标准株和1株牛带绦虫标准株的成节或孕节节片,提取总DNA,随机选择12条引物进行RAPD分析,SPSS软件构建不同地理株系统发育树。结果12条随机引物共扩增片段134个(以相同bp数为依据),单条引物扩增片段范围为3～22个,平均扩增片段11.17个。P1～6、P9和P11可扩增出特异条带。亲缘关系树状图显示,怒江1～4号(NJ1～4)、迪庆1～3号(DQ1～3)、大理2～3号(DL2～3)与亚洲带绦虫都匀株标准(BZ1)和亚洲带绦虫大理株(BZ3)汇为一支,属于亚洲带绦虫;临沧1号(LC1)、楚雄1号(CX1)、西双版纳1号(BN1)和牛带标准株(BZ2)汇为一支,属于传统牛带绦虫;2支结合后与DL1结合。结论云南省存在3种带绦虫的流行。RAPD可以用于带绦虫鉴定,同时也为亚洲带绦虫的具体分类定位提供参考依据。

Objective To make molecular identification and analysis of 14 isolates of taenia from NUJIANG, DIQING,DALI, LINCANG,CHUXIONG and XISHUANGBANNA in Yunnan Province. Methods After Morphological identification,genomic DNA was extracted from the segments of ddults or the scolex fragment of 13 isolates of T. saginata , 1 isolate of T. solium,2 standard strains of T. asiatica and 1 standard strains of T. saginata,RAPD were carried out with 12 random primers. Different geography system growth tree was constructed by means of SPSS software. Results 134 DNA fragments were amplified by 12 primers. The number of DNA fragments amplified by single primer was between 3 and 22,and the average was 11.17. Electrophoresis of P1-6,P9 and Pll have specific DNA fragments. The rescaled distance cluster combination demonstrated that NJ1-4, DQ1-3 and DL2-3 occupied one branch with BZ1 and BZ3,which belongs to T. asiatica. LC1,BN1 and CX1 occupied one branch with BZ2,which belongs to T. saginata . Two hranchs occupied with DL1. Conclusion There are three kinds of taenia in the area of Yunnan,and RAPD can be used in the identification of the taenia and may provide the basis for the molecular classifi- cation of T. asiatica.