交叉引物等温扩增技术在恶性疟疾快速检测中的应用  

Application of cross priming amplification in rapid detection of falciparum malaria

作　　者：祁军[1] 于智睿[1] 詹曦菁[1] 黄吉城[2] 李智慧[1] 

机构地区：[1]天津出入境检验检疫局,天津300457 [2]广东出入境检验检疫局

出　　处：《中国媒介生物学及控制杂志》2013年第3期204-207,共4页Chinese Journal of Vector Biology and Control

基　　金：国家质量监督检验检疫总局项目(2010IK221)~~

摘　　要：目的建立一种快速、敏感、特异检测恶性疟原虫的交叉引物等温扩增(cross priming amplification,CPA)技术。方法根据疟原虫的18SrRNA基因保守序列,设计CPA引物和探针。分别用CPA法、吉氏染色镜检法检测78份发热患者血样,验证CPA法特异性和灵敏性。结果采用CPA法对不同病原体核酸的检测结果显示,38份含有恶性疟原虫血样中检出37例阳性,其他病原体检测结果均为阴性,证明其特异性良好;该方法检测的灵敏度可达102拷贝/μl;与镜检法比较,其检测的灵敏度为97.37%,特异性为100%,准确度为98.72%。结论用于检测恶性疟原虫的CPA检测体系具有简便、快速、灵敏、特异的优点,可用于临床、预防和出入境口岸现场疟疾的快速检测。Objective To develop cross priming amplification （CPA） technology for rapid, sensitive, and specific detection of Plasmodium falciparum. Methods The CAP primers and probes were designed based on the conserved 18S rRNA sequence of P. falciparum. The blood samples of 78 patients with fever were examined by CPA and Giemsa staining microscopy, and the specificity and sensitivity of CPA method were evaluated. Results P. falciparum was detected by CPA method in 37 of 38 P. falciparum containing blood samples, and other pathogens were not detected by this method, suggesting that CPA method has high specificity for P. falciparum. The sensitivity of CPA method was 10 2 copies/μl. The specificity, sensitivity, and accuracy of CPA method were 97.37%, 100%, and 98.72%, respectively, as compared with those of microscopic examination. Conclusion CPA assay is a simple, rapid, sensitive, and specific method for detecting P. falciparum in the clinical setting and at entry exit ports.

关 键 词：交叉引物等温扩增 恶性疟疾 疟原虫 

分 类 号：R531.3[医药卫生—内科学]