套式PCR扩增特定SSrRNA基因片段诊断间日疟及混合感染的研究  

DETECTION OF PLASMODIUM VIVAX AND MIXED INFECTION BY AMPLIFYING SPECIFIC SSrRNA GENE WITH NESTED POLYMERASE CHAIN REACTION

作　　者：诸欣平[1,2,3] 李明[1,2,3] 董洁[1,2,3] 周蕾 王学忠[1,2,3] 王凤云 

机构地区：[1]首都医科大学 [2]第一军医大学热带医学研究所 [3]云南省疟疾防治研究所

出　　处：《中国寄生虫病防治杂志》1998年第2期92-95,共4页Chinese Journal of Parasitic Disease Control

基　　金：北京市教委科技发展基金;全军医药卫生科研基金

摘　　要：根据间日疟原虫（Ｐ．ｖ．）小亚单位核糖体核糖核酸（ＳＳｒＲＮＡ）基因序列，设计并合成两对特异引物，采用套式聚合酶链反应（ＰＣＲ）技术，扩增出一条长１２０个碱基对（ｂｐ）的间日疟原虫ＳＳｒＲＮＡ基因特定片段，并用于云南间日疟患者的血样检测。结果表明：该系统特异性强，除间日疟原虫外，恶性疟原虫（Ｐ．ｆ．）、三日疟原虫（Ｐ．ｍ．）及正常人血ＤＮＡ样本均无此扩增带出现。该系统检测原虫的敏感度为０．１个疟原虫／ｌμｌ血，大大高于常规镜检。在进行间日疟检测中，该系统与镜检的阳性符合率为１００％，更重要的是发现镜检漏诊的４例Ｐ．ｖ．和Ｐ．ｆ．的混合感染病例。鉴于该系统具有特异、敏感、稳定、简便的特点和鉴别疟原虫种类、判定混合感染等优点，故对疟疾的诊断。Two pairs of oligonucleotide primers were designed and synthesized based on small subunit ribosomal RNA gene of plasmodium vivax , and blood samples obtained from malaria vivax patients in Yunnan were analyzed by nested polymerase chain reaction (PCR ). The results showed that a 120 base pairs(bp) DNA fragment of P.vivax was amplified. No amplification was observed with P. falciparum , P. malariae and human DNA. The nested PCR could detect parasitemia to an extent as low as 0.1 parasite/μl. The positive concordance rate between nested PCR and microscopy was 100% in detecting P.vivax . In addition, 4 cases of mixed plasmodium infection were found by PCR. It is suggested that this method is specific, sensitive, stable and has a great potential in identifying different plasmodium species and detecting mixed infections. Thus, PCR assay provides a powerful tool in study of malaria diagnosis and epidemiology.

关 键 词：疟疾 SSRRNA PCR 间日疟原虫 混合感染 诊断 

分 类 号：R531.304[医药卫生—内科学]