套式PCR扩增特定SSUrDNA片段诊断恶性疟的研究  

STUDIES ON DIAGNOSIS OF FALCIPARUM MALARIA BASED ON AMPLIFYING SPECIFIC SSUrDNA FRAGMENT WITH NESTED PCR

作　　者：万磊[1] 陈培霞[1] 薛采芳[1] 姜绍谆[2] 

机构地区：[1]第四军医大学基础部寄生虫学教研室,西安710032 [2]第四军医大学基础部微生物学教研室,西安710032

出　　处：《中国寄生虫学与寄生虫病杂志》1995年第3期174-177,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金!39070753

摘　　要：设计2对针对恶性疟原虫（P．f.）小亚单位核糖体核糖核酸基因（SSUrDNA）的特异引物，采用双温度点套式聚合酶链式反应（PCR）技术，从用煮沸法快速萃取的样本DNA中，扩增P．f.SSUrDNA片段，进行恶性疟原虫的检测。结果表明，该系统扩增样本DNA片段大小恒定，经限制性酶切进一步分析，证实均为P．f.SSUrDNA目的片段，其检测原虫的灵敏度为0．8×10-6，较常规镜检敏感，并具有鉴别红内期早期阶段疟原虫种类和确认有无混合感染的潜力。因而认为该系统是灵敏、准确、简易、快速的疟疾诊断新方法，值得推广使用。Two pairs of primers specific to small subunit ribosomal DNA of Plasmodium falciparum were designed and the expected SSUrDNA fragment was amplified for detecting P.falciparum infection with double-ternperature-nested polymerase chain reaction using DNA prepared by boiling method. The results showed that the nested PCR could amplify a constant size of desired SSUrDNA fragment of P. falciparum which was further confirmed by digestion of restrlction endonuclease and could detect parasitemia level of 0. 8 × 10-6. It has great potentials for identifying Plasmodium species in ring form of erythrocytic stage and detecting mixed Plasmodium infections. Therefore, it is suggested that this method is sensitive, accurate, simple and rapid in detecting Plasmodium falciparum in blood samples for malaria diagnosis.

关 键 词：疟原虫 疟疾 聚合酶链反应 基因诊断 

分 类 号：R531.304[医药卫生—内科学]