基因扩增检测四种感染人的疟原虫种类、数量的方法  

Methods for Detecting Four Species of Human-infected Plasmodia and Their Amount with Gene Amplication

作　　者：陶玉滨[1] 许勇臣[1] 杨利[1] 

机构地区：[1]中国人民解放军第211医院,黑龙江哈尔滨150086

出　　处：《实用预防医学》2008年第4期1044-1047,共4页Practical Preventive Medicine

摘　　要：目的建立对四种感染人的疟原虫种类、数量进行基因检测的方法。方法根据恶性疟、三日疟、卵形疟、间日疟的18SrRNA基因序列,设计属、种特异性引物和TaqMan探针,用荧光定量扩增反应确定标本中的基因拷贝数,用电泳区别各种疟原虫,并对2份疟疾血样进行检测。结果建立的检测方法对疟疾血样进行检测,荧光定量PCR具有良好的反应性,通过电泳能区分检测标本中的疟原虫株。结论建立的方法能够特异而灵敏地检测出标本中疟原虫的种类、数量,适合疟疾防治检测的需要。Objective To develop a series of detecting methods for the quantification and quantitation of four species of human- host plasmodia. Methods Genus - specific primers, species - specific primers, and a TaqMan probe were designed in accordance with the 18S rRNA gene of Plasmodium falciparum, Plasmodiun vivax, Plasmodium malariae and Plasmodiun ovale. Recombinant plasmid of species was constructed. A fluro- quantitative polymerase chain reaction （PCR） was developed to determine the copies and the electrophoresis for the identification. Two positive samples were tested. Results Specific amplification was achieved with the primers and the excellent Iinearity was exhibited in the fluro - quantative PCR which had been established with the standard of recombinant plasmid. Conclusions The detecting system is established successfully with high specificity and sensitivity, which satisfies for the prevention and control of malaria.

关 键 词：聚合酶链反应 定性 定量 疟原虫 18S RRNA基因 

分 类 号：R382.31[医药卫生—医学寄生虫学]