套式PCR法检测间日疟原虫的研究  

Study on Detection of Plasmodium uiuax in Blood Sample fromMalaria-Endemic Area by the Nested Polymerase Chain Reaction

作　　者：孙明林[1] 陈培霞[1] 薛采芳[1] 万磊[1] 刘忠湘[1] 

机构地区：[1]第四军医大学寄生虫学教研室

出　　处：《地方病通报》1996年第1期17-20,共4页Endemic Diseases Bulletin

基　　金：国家自然科学基金

摘　　要：以滤纸采集间日疟原虫患者血样，点沸法萃取ＤＮＡ模板，用间日疟原虫ＳＳＵｒＤＮＡ特异序列为内外引物，进行双温度点套式ＰＣＲ扩增，其检测原虫率水平为０．８４×１０￣（－６），特异性强。检测云南疟区发热患者血样１６６份，检出率为６３．９％，显著高于镜检法的４８．２％。以镜检法为参照，本法的敏感性为９７％，特异性为６１％，阳性预期值和阴性预期值分别为６５％和９７％，表明是检测间日疟原虫感染的一个良好手段，可作为疟区流行病学监测、药物和疫苗效果考核及供血员筛选的有效工具。The nested polymerase chain reaction based on amplifying Plasmodium uiuax ( P.u)special SSUrDNA fragment was applied to detect P. u from drled-blood-spotted filter paper.The detecting level withthis method was less than 0.84×10(-5)in parasitemia(<10 parstites/μl whole blood).116 samples were col-lected from fevet patients presenting at clinics in Yunnan malaria-endemic area and analyzed both by nested PCR method and by conventional microscopy.Out of 116,69 were positive both by nested PCR and by mi -croscopy,while 58 were negative by both niethods.The postive rate by nested PCR(63.9%) was highlystastically significant than that by microscopy(42.8%).Compared with microscopic examination,the sensi -tivity and the specificity by nested PCR was 97%and 61%respectively.The results showed that this nested PCR method should be suitable for detection of P.u with low parastemia and may prove to be a vauable toolfor large-scale epidemiologic study,follow-up of drug treatment and vaccine trial,and blood-donor screening.

关 键 词：疟疾 间日疟原虫 聚合酶链反应 基因诊断 

分 类 号：R531.304[医药卫生—内科学]