摘要
目的建立一种简便快速、能同时检测恶性疟和间日疟的核酸检测方法。方法针对两种疟原虫18SrRNA基因设计2对(3条引物),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多重PCR。并进行最低检测限确定和临床标本检测,以镜检法为金标准分析灵敏度和特异度等指标。结果该方法可扩增出431bp(恶性疟原虫)和341bp(间日疟原虫)基因片段,最低检测限为102copies/反应,检测临床标本的结果与镜检法无差别(P>0.05),敏感度为93.55%,特异度为70.83%,阳性预测值为89.23%,阴性预测值为80.95%。结论所建立的多重PCR方法可快速检测疟疾感染并鉴别分型,灵敏度高,值得推广。
A multiplex PCR to detect Plasmodium falciparum （P. falciparum） and 191asmodiurn vivax （P. vivax） was established simply and rapidly in our study. Two pairs of primers （3 strips） were designed according to the sequences of the 18S rRNA of the two kinds of Plasmodium. The reacting system was optimized by analysis on the different primer concen- tration and annealing temperature. The patients＇ blood samples were tested with the optimized multiplex PCR, and the limit of detection of this assay was estimated. The sensitivity and specificity were calculated using microscopy as the gold standard. The sizes of amplification products of multiplex PCR amplifying genomic DNA of P. falciparum and P. vivax were 431 bp and 341 bp, respectively. The limit of detection of this assay was 102 copies/reaction. The clinical sensitivity was 93.55 % and the specificity was 70.83%. The positive predictive value （PPV） was 89.23% and the negative predictive value （NPV） was 80.95%. So we concluded that the multiplex PCR with high sensitivity could detect and identify malaria rapidly, which is suit- able for primary Centers for Disease Prevention and Control.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2014年第3期288-291,共4页
Chinese Journal of Zoonoses
基金
中南大学中央高校基本科研业务费专项资金资助(No.2013zzts299)
长沙市科技计划项目(No.K1205032-31)
湖南省科技计划项目(No.2012SK3301)
国家质检总局科技计划项目(No.2013IK224)~~
关键词
疟疾
恶性疟原虫
间日疟原虫
多重PCR
malaria
Plasmodium falciparum
Plasmodium vivax
multiplex PCR
