套式PCR检测滤纸干血滴中恶性疟原虫的研究  

DETECTION OF PLASMODIUM FALCIPARUM FROM DRIED BLOOD SPOTTED FILTER PAPER BY NESTED PCR

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：《中国寄生虫学与寄生虫病杂志》1996年第3期197-200,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金

摘　　要：目的：建立简易、实用的套式ＰＣＲ检测恶性疟原虫（Ｐ．ｆ．）的方法。方法：以小亚单位核糖体核糖核酸基因（ＳＳＵｒＤＮＡ）为目的片段的双温度点套式ＰＣＲ扩增滤纸干血滴中Ｐ．ｆ．ＤＮＡ，并将结果与镜检法进行比较研究。结果：１４３例标本中两法均阳性和均阴性分别有５５份和５１份，镜检阴性而ＰＣＲ阳性者３４份，镜检阳性而ＰＣＲ法阴性３份。套式ＰＣＲ总阳性率为６２％，显著高于镜检法的４１％，两者符合率为７４％。结论：本法简便、快速，敏感性高、特异性强，具有一定的现场应用价值。AIM:To establish a simple and practical system of nested PCR to detect Plasmodium falciparum (P.f.). METHOD: Comparing with the conventional microscopy method, the two temperature point nested PCR was applied to detect P.f. from dried blood spotted filter paper with two sets of oligonucleotide primer pairs specific to P.f. small subunit ribosomal RNA gene (SSUrDNA). RESULTS: This method was successful in detecting parasitemia of less than 1.3×10 -7 (approx. 1 parasite/2μl whole blood). 143 samples were collected from febrile patients at clinics in Yunnan malaria endemic area and were analysed both by nested PCR method and by microscopy. Of these, 55 were positive both by microscopy and by nested PCR while 51 were negative by both methods. Nested PCR detected 34 samples that were negative for P.f. by microscopy, and failed to detect 3 samples identified as positive by microscopy. The positive rate of the nested PCR (62%) was found highly statistically significant than that of microscopy method (41%). The concordance rate between these two diagnostic methods was 74%. CONCLUSION: The ease of collection and transport of filter paper specimens combined with the sensitive and specific detection of P.f. by nested PCR suggest that this method might be a valuable tool for epidemiological study of falciparum malaria.

关 键 词：恶性 疟原虫 滤纸干血滴 检测 

分 类 号：R382.31[医药卫生—医学寄生虫学]