逆转录聚合酶链反应用于检测恶性疟原虫的初步研究  

A Preliminary Study on Detecting Plasmodium Falciparum Using Reverse Transcriptase polymerase Chain Reaction

作　　者：胡锡敏[1] 王善青[1] 金玉明[1] 童重锦[1] 华德[1] 陈冬燕[1] 曾文[1] 

机构地区：[1]海南省热带病防治研究所,海口570203

出　　处：《中国公共卫生》2001年第5期396-397,共2页Chinese Journal of Public Health

基　　金：海南省自然科学基金资助项目

摘　　要：目的　建立逆转录聚合酶链反应系统 ,用于检测恶性疟原虫感染。方法　以恶性疟原虫小亚单位核糖体核糖核酸基因为靶片段 ,设计一对特异引物 ,分别采用RT -PCR技术和PCR技术进行检测恶性疟原虫血样并比较两者检测的敏感性。结果　从培养的恶性疟血样中扩增出 35 9bp特定扩增带 ,而间日疟和健康人血样均未见扩增带。经限制性内切酶分析证实扩增产物为目的片段。检测系统初步显示可检出恶性疟血样中 0 34个疟原虫所含的RNA模板。以RNA为检测对象的RT -PCR肉眼可见条带的检测水平较以DNA为检测对象的PCR扩增高。结论　RT -PCR检测恶性疟原虫方法具有敏感性高、特异性强的特点 。Objective To develop a new method based on reverse transcriptase polymerase chain reaction(RT-PCR) for detecting plasmodium falciparum(p.f).Methods Primers were designed according to the asexual stage p.f SSUrRNA gene.RT-CPCR system was established and applied to detect p.f.Results A 359bp fragment from cultered p.f was amplified,while the extracted RNA from P.vivas and the normal blood sample could not.The amlified fragments was confirmed by means of restriction endonuclease digestion.As little as RNA of 0.34 paraste was sufficient for detection by RT-PCR assay.The detection sensitivity of RT-PCCR could by higer than that of PCR.Conclusion The RT-PCR assay was specific and sensitive in the detection of p.f.

关 键 词：恶性疟原虫 逆转录聚合酶链反应 诊断 

分 类 号：R382.31[医药卫生—医学寄生虫学]