摘要
为确立一种快速、敏感、特异的疟原虫检测方法,根据红内期疟原虫SSUrRNA基因序列,设计合成引物3条,采用微量全血和滤纸干血滴标本快速制备疟原虫DNA模板的9种方法,进行聚合酶链反应(PCR)并比较。结果显示有5种方法的实验效果最佳,其中,全血PCR仪预热及滤纸干血滴Chelex煮沸两法操作简单、所需试剂较少。采用该两法对深圳地区202份和湖北地区16份镜检阳性的全血和深圳地区129份滤纸干血滴标本进行检测,结果与镜检的符合率分别为98.5%和99.2%。表明本研究筛选出的两种方法制备DNA模板的PCR检测体系,适用于临床快速准确诊断间日疟、恶性疟或两者混合感染,尤其适用于流行病学调查,值得推广。
To establish a rapid, sensitive and specific method to detect malaria parasite, three primers were designed according to the SSUrRNA gene sequences of malaria parasite. And nine methods of rapidly preparing DNA from micro blood samples were used for polymerase chain reaction(PCR) and compared with each other. The results showed that five methods were the best. Among these methods, both were selected to detect 218 blood samples and 129 dried blood spot samples of patients diagnosed by microscopiests. The results indicated that 215 of 218(98.5%) whole blood samples and 128 of 129(99.2%) dried blood spot samples were positive by the PCR amplification and 10 normal blood samples were all negative.
出处
《中国寄生虫病防治杂志》
CSCD
1998年第4期260-263,共4页
Chinese Journal of Parasitic Disease Control