RAPD技术分析嗜人按蚊种型方法的建立

目的：建立用RAPD－PCR技术鉴别中国不同纬度地区嗜人按蚊，特别是海南岛嗜人按蚊与大陆嗜人按蚊间有无种型差异的新方法。方法：收集海南，四川，江苏三地嗜人按蚊标本并传代培养，优化蚊基因组的DNA的提取纯化及RAPD－PCR扩增和产物检测的条件，筛选5条RAPD引物，结果：筛选出1条适合三地嗜人按蚊的引物并制定出了稳定的RAPD图谱，建立了RAPD－PCR分析嗜人按蚊种型的方法。结论：不同纬度三地嗜人按蚊大部分谱带是相同的，但它们的谱带数不一定相同，且少数谱带是完全不同的，初步表明我国嗜人按蚊存在一定的种型差异，从而为解释海南嗜人按蚊在生态习性和传疟作用方面为何与大陆嗜人按蚊不同从分子水平提供了一定的科学依据。

Establishment of random amplified polymorphic DNA polymerase chain reaction (RAPD- PCR) method in analysis of Anopheles anthopophagus

ve To establish a new method based on random amplified polymorphic DNA polymerase chain reaction (RAPD - PCR) for identifying if there is the species' difference among Anopheles anthropophagus of different latitudes, especially between Hainan Island and the inner land in China. Methods The samples of Anophes anthropophagus were collected from Hainan Island, Sichuan Province and Jiangsu Province, and cultured in the laboratory. To improve the methods of extraction and purification of mosquito genomic DNA, amplification with RAPD - PCR technique and detection of amplified fragments repeatedly, and screen primers. Results A new method of RAPD - PCR was established for species differentiation of Anopheles anthropophagus , one primer was screened out from 5 RAPD single primers and the stable RAPD maps were obtained. Conclusion Most of the amplfied fragments were common for the three areas' Anopheles anthropophagus, but some fragments were unique to one area's Anophes anthropophagus. It indicated there was the species difference of Anophes anthropophagus in China, and had provided the scientific evidence of molecular biology to explain why the ecological habits and malaria - transmitting roles of Anopheles anthropophagus in Hainan were different from that in the inner land.