【摘要】 为建立能检测蚊体内恶性疟原虫的聚合酶链反应技术 ,根据恶性疟原虫 CSP基因序列 ,设计合成 1对引物 ,以 Chelex- 10 0煮沸法制备 DNA模板 ,采用 PCR方法 ,恶性疟原虫子孢子模板预期被扩增出 2 45 bp的 DNA条带 ,并将该法与蚊虫解剖镜检子孢子方法相比较。结果经人工感染恶性疟原虫的 31只大劣按蚊中 14只 PCR阳性 ,被扩增出预期大小的 DNA片段 ,其余 17只 PCR阴性 ;以该技术检测野外捕捉的 98只按蚊 ,结果均为阴性。 PCR法与解剖镜检法相比较 ,检测结果完全符合 ;相当于 1/ 10个阳性蚊子的模板即可满足 PCR检测。该检测体系灵敏、特异 ,对于蚊体内恶性疟原虫的检测具有一定的应用价值更多还原

【Abstract】 To establish a PCRbased method to detect Plasmodium falciparum sporozoites in mosquitoes. A pair of primers based on the P. falciparum circumsporozoite protein (CSP) gene sequence was designed, and a 245 bp DNA fragment was amplified from the extracts of mosquitoes infected with P. falciparum by PCR. The PCRbased method was also compared with traditional microscopical method. The results showed that 14 of 31 mosquitoes infected with P. falciparum were positive by the PCRbased method and microscopical method, and 98 mosquitoes collected wildly were all negative by both method. The PCRbased method was sensitive and specific in detecting P. falciparum sporozoites in mosquitoes.更多还原