逆转录聚合酶链反应检测和鉴定疟原虫的研究  

Detection and identification of plasmodia by mean of reverse transcriptase polymerase chain reaction

作　　者：胡锡敏[1] 林世干[1] 陈冬燕[1] 崔宜庆[1] 童重锦[1] 王善青[1] 

机构地区：[1]海南省疾病预防控制中心

出　　处：《中国人兽共患病杂志》2004年第10期894-896,共3页Chinese Journal of Zoonoses

基　　金：海南省自然科学基金资助项目(No39713)

摘　　要：在同一反应体系中建立能同时检测和鉴定恶性疟、间日疟及混合感染的逆转录聚合酶链反应(RT-PCR)检测方法,并用于现场采集血样检测。先以恶性疟原虫小亚单位核糖体核糖核酸基因为靶片段,设计1对针对恶性疟原虫的特异性引物,建立RT-PCR检测恶性疟原虫方法,在此基础上,增加1条针对间日疟原虫的种特异性引物,使在同一反应体系中,能够同时扩增恶性疟原虫和间日疟原虫的特异片段。恶性疟原虫和间日疟原虫模板在同一反应体系中分别被扩增出预期大小为359bp和449bp特定扩增带,健康人血样和空白对照均未见扩增带。用建立的RT-PCR检测方法对现场采集128份血样进行检测,与镜检的阳性符合率为9609%,14份镜检为阴性的发热病人全血标本中2份RT-PCR检测为间日疟阳性。RT-PCR检测疟原虫方法具有敏感性高、特异性强的特点,对疟疾诊断、镜检质量控制和流行病学研究具有较大的实用价值。To develop a method to detect the presence of Plasmodium falciparum and Plasmodium vivax by means of reverse transcriptase polymerase chain reaction (RT PCR),a common upper primer and two species specific lower primers of P.falciparium and P.vivax were designed according to the sequences of the small subunit ribosomal RNA (SSUrRNA) fragment of these two species of plasmodia.RT PCR system was established and used to detect the presence of plasmodia in the blood samples in field survey.It was found that the DNA fragments of about 359 bp and 449 bp in length were successfully amplified by RT PCR from the genomic DNA of plasmodia,but no fragment was obtained from that of blood of healthy persons.Of 128 samples detected by this method,123 samples were coincident with the results of microscopic examinations in which two blood sample of P.vivax infection that was not to be positive by microscopic examination were detected by RT PCR system.Thus,this RT PCR system might be useful for the diagnosis of malaria,quality control for the microscopic examination and epidemiological studies.

关 键 词：恶性疟原虫 间日疟原虫 逆转录聚合酶链反应 检测 

分 类 号：R531.3[医药卫生—内科学]