国境口岸疟疾快速联合检测方法应用评估

    方筠，韩晓辉，贾哲甫，叶魏，章琪，张晓航，王健，陆晔

    上海国际旅行卫生保健中心，上海 200335

    摘要：目的 对疟疾快速检测方法的联合应用进行评价，探索适用于口岸的疟疾快速检测方法。方法选取来源于中国科学院寄生虫病研究所的疟原虫镜检阳性滤纸血样本，其中30例恶性疟，28例间日疟和20例正常人对照样本；同时使用检测疟疾的实时荧光PCR方法、交叉引物恒温扩增方法、胶体金技术对其进行检测；分析各方法检测的敏感性和特异性。结果实时荧光PCR法检测恶性疟原虫的灵敏度高于交叉引物恒温扩增法和胶体金法（P值分别为0.019、0.043，P＜0.05)）；对低原虫密度的恶性疟样本（原虫密度10~100及101~1000个/μl），实时荧光PCR法检测的灵敏度较高，交叉引物恒温扩增法和胶体金法对于低原虫密度的恶性疟样本检测灵敏度显著低于PCR法；交叉引物恒温扩增和胶体金法联合检测在恶性疟各原虫密度下的灵敏度均高于使用单一方法的灵敏度，与实时荧光PCR法的灵敏度比较，差异无统计学意义（各原虫密度下，两者比较的P值分别为0.29、0.302、1.000、1.000，P＞0.05）。实时荧光PCR法对间日疟样本检测灵敏度较高，交叉引物恒温扩增和胶体金法的灵敏度显著低于实时荧光PCR法，两者共检能提高检测的灵敏度。结论交叉引物恒温扩增法和胶体金法联合检测，可以较好地筛选出各种原虫密度的感染者，该联合检测方法的灵敏度和特异性与实时荧光PCR法相当，适合国境口岸对疟疾现场快速检测的要求。

    关键词：疟疾；实时荧光PCR；交叉引物恒温扩增；胶体金；应用；评估；国境口岸

    中图分类号：R183.5　文献标识码：B

    Evaluation on application of the malariarapid screening system at ports

    FANG Yun, HAN Xiao-hui, JIA Zhe-fu, YE Wei,ZHANG Qi, ZHANG Xiao-hang, WANG Jian, LU Ye

    Shanghai International Travel HealthcareCenter,Shanghai 200335,China

    Abstract:   Objective  To establish and evaluate a rapid system for screening malaria atports and basic clinical laboratory.  Methods   Using three kinds of by real time florescentpolymerase chain reaction (PCR), Crossing Priming Amplification(CPA), Colloidal gold technique to detect malaria blood samples(30cases of Plasmodium falciparum, 28 cases of Plasmodium vivax and 20cases of normal control) on the filter paper from Institute ofParasitic Diseases in Shanghai. The specificity and sensitivity ofall the methods were evaluated.   Results   Thesensitivity of real time florescent PCR was high than CPA andColloidal gold technique in Plasmodium falciparum samples, and evenmore significant in the low parasite density sample (parasitedensity 10-100,101-1 000/μl). The sensitivity of combining use ofCPA and Colloidal gold technique was high than single method in allparasite density. And it was not different from PCR in statistics.A similar phenomenon was observed in Plasmodium vivax.Conclusion   Combining use of CPA and Colloidal goldtechnique rapid diagnostic devices is a better system to screenmalaria infect persons in all parasite density. The sensitivity andspecificity of the system are equal to PCR, and it could be used asa screen method in preventing and controlling malaria at ports andbasic clinical laboratory.

    Key words:   Malaria; Real timeflorescent PCR; Crossing priming amplification; Colloidal goldtechnique

    《中国国境卫生检疫杂志》