疟疾的实时荧光PCR快速检测方法  

Real-time fluorescent PCR method for rapid detection of malaria

作　　者：师永霞[1] 苏锦坤[1] 洪烨[1] 李小波[1] 郑夔[1] 黄吉城[1] 幸芦琴[1] 相大鹏[1] 郭波旋[1] 

机构地区：[1]广东出入境检验检疫局检验检疫技术中心,广州510700

出　　处：《中国卫生检验杂志》2011年第3期625-627,共3页Chinese Journal of Health Laboratory Technology

基　　金：广东检验检疫局科技计划项目(2007GDK32)

摘　　要：目的:建立疟疾的实时荧光PCR检测方法并用于疟疾的快速检测。方法:设计了疟原虫通用型实时荧光PCR检测的引物和探针,建立了疟疾的实时荧光PCR检测方法。对40份疟疾镜检阳性样本和20份镜检阴性样本进行实时荧光PCR检测,并和巢式PCR方法检测结果进行了比较。结果:疟疾实时荧光PCR检测速度快,20分钟即可完成。40份镜检阳性样本的荧光PCR检测均为阳性,2份样本的巢式PCR结果为阴性;20份镜检阴性样本中有3份样本为实时荧光PCR阳性,其余17份样本为实时荧光PCR阴性,与疟疾巢式PCR检测结果相同。结论:该实时荧光PCR方法检测速度快,特异性强,准确性高,阳性率高,不易漏检,适用于疟疾的快速检验,这对防止该病在国境口岸的传入提供了技术依据。Objective：To establish real-time fluorescent PCR detection method for rapid detection of malaria.Methods：Universal primers and probes were designed.Forty positive samples and twenty negative samples tested by microscopic examination were detected Plasmodium by real-time fluorescent PCR.Their results were compared with that of nested PCR.Results：Real-time fluorescence PCR detection of malaria was fast and the reaction time only took 20 minutes.All of forty microscopy-positive samples were positive for fluorescent PCR detection,and two samples were negative by nested PCR.Three of twenty microscopy-negative samples were positive for real-time fluorescence PCR and the remaining seventeen samples were negative,which was the same as results of nested PCR detection.Conclusion：The real-time fluorescent PCR is suitable for rapid detection of malaria because of its fast speed,strong specificity,high accuracy and low missing rate.It provides a technical basis to prevent disease from spreading into our country in the border.

关 键 词：疟疾 小亚单位核糖体核糖核酸 实时荧光PCR 巢式PCR 

分 类 号：R531.3[医药卫生—内科学]