SYBR Green实时PCR检测恶性疟原虫和间日疟原虫的临床检验技术研究  

Research on SYBR Green based real time PCR for detection of Plasmodium falciparum and Plasmodium vivax

作　　者：李欢[1,2] 王晓春[1] 

机构地区：[1]湖南省妇幼保健院检验科,长沙410005 [2]中南大学湘雅医学院医学检验系,长沙410013

出　　处：《国际医学寄生虫病杂志》2015年第5期255-260,共6页International JOurnal of Medical Parasitic Diseases

摘　　要：目的：建立鉴别恶性疟和间日疟的SYBR Green实时PCR检测方法。方法针对恶性疟原虫和间日疟原虫18S rRNA基因设计引物，优化引物浓度与退火温度，建立可扩增出两种疟原虫基因片段的SYBR Green 实时PCR，并进行54例临床标本检测，其中恶性疟32例、间日疟22例。以镜检法为金标准分析敏感度和特异度等指标。结果该方法可扩增出恶性疟原虫和间日疟原虫18S rRNA 基因片段，并能检出混合感染，特异性好。该方法检测恶性疟原虫或间日疟原虫的敏感度为97.30%（36/37），特异度为5.88%（1/17）；检测恶性疟原虫，敏感度为87.50%（21/24），特异度为63.33%（19/30）；检测间日疟原虫，敏感度为69.23%（9/13），特异度为68.29%（28/41）。结论所建立的SYBR Green 实时PCR方法能较准确地诊断疟疾并鉴别虫种，敏感度高，在混合感染的诊断方面具有优越性。Objective To establish a SYBR Green based real time PCR to detect Plasmodium falci-parum and Plasmodium vivax. Methods The primers were designed according to the sequences of 18S rRNA of the two species of Plasmodium. The reaction system was optimized through different primer concen-trations and annealing temperatures. Fifty-four patients’ blood samples （32 with P. falciparum and 22 with P. vivax） were tested with the optimized SYBR Green real time PCR. The sensitivity and specificity were evalu-ated by microscopic examination as the gold standard. Results The sensitivity of detection on P. falci-parum or P. vivax malaria was 97.30%（36/37） and the specificity was 5.88%（1/17）. For detection on P. fal-ciparum,the sensitivity was 87.50%（21/24） and the specificity was 63.33%（19/30）. For detection on P. vi-vax,the sensitivity was 69.23%（9/13） and the specificity was 68.29%（28/41）. Conclusion The SYBR Green real time PCR could detect and identify malaria with high sensitivity and fair specificity.

关 键 词：疟疾 恶性疟原虫 间日疟原虫 SYBR Green实时PCR 

分 类 号：R379.4[医药卫生—病原生物学]