一种简易疟原虫PCR模板制备方法

目的:寻找一种高效、简便、快速疟原虫PCR模板制备方法。方法:收集2010年-2011年自非洲、东南亚等疟疾流行区回国人员中疟疾现症患者(显微镜检查阳性),采滤纸血,用蒸馏水直接溶血法和Chel-ex-100法分别制备疟原虫PCR模板,采用巢式PCR技术检测疟原虫ss RNA基因,并对两法结果进行比较。结果:分别用蒸馏水直接溶血法和Chelex-100法制备疟原虫PCR模板32个,其中恶性疟原虫26个,间日疟原虫6个,直接溶血法巢式PCR检测全部出现特异性扩增带,而Chelex-100法均未出现扩增带。结论:蒸馏水直接溶血法是一种较为理想制备疟原虫PCR模板的方法。

A simple method for preparing template of Plasmodium for PCR

Objective:To explore a simple,rapid and efficient method for preparing template of Plasmodium for polymerase chain reaction(PCR).Methods:Dried blood spot samples of malaria patients from Africa and Southeast Asia who were diagnosed as positive by microscopy were collected on filter paper from 2010 to 2011.Two methods for preparing template of Plasmodium,direct hemolysis method by distilled water and Chelex-100 method,were used respectively for nested PCR to detect ss RNA gene of Plasmodium and compared with each other.Results:Thirty-two templates of Plasmodium with twenty-six of P.falciparum and six of P.vivax were prepared by both methods.Nested PCR with template prepared by direct hemolysis method showed all the samples produced specific amplification bands.But no bands were detected by nested PCR with templates prepared by Chelex-100 method.Conclusion:Direct hemolysis method was perfect to prepare templates of Plasmodium for nested PCR.