一种简易疟原虫PCR模板制备方法  

A simple method for preparing template of Plasmodium for PCR

作　　者：周水茂[1] 刘勇[1] 陈智[1] 吴凯[1] 杨燕[1] 王重新[1] 徐明星[1] 

机构地区：[1]湖北省武汉市疾病预防控制中心,武汉430015

出　　处：《中国卫生检验杂志》2012年第10期2385-2387,共3页Chinese Journal of Health Laboratory Technology

摘　　要：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。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 South- east Asia who were diagnosed as positive by microscopy were collected on filter paper from 2010 to 2011. Two meth- ods 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. Re- sults ： 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 spe- cific amplification bands. But no bands were detected by nested PCR with templates prepared by Chelex - 100 meth- od. Conclusion： Direct hemolysis method was perfect to prepare templates of Plasmodium for nested PCR.

关 键 词：疟原虫 PCR模板 直接溶血法 Chelex-100 

分 类 号：R531.3[医药卫生—内科学]