应用量子点及磁微粒技术快速检测阴道毛滴虫抗原  

Rapid detection of Trichomonas vaginalis antigens by quantum dots and magnetic beads

作　　者：李蒙[1] 蒲晓允[1] 李玲[1] 余娟春[1] 

机构地区：[1]第三军医大学新桥医院检验科,重庆400037

出　　处：《第三军医大学学报》2009年第8期740-743,共4页Journal of Third Military Medical University

基　　金：重庆市人口和计划生育委员会资助项目(06-2-04)~~

摘　　要：目的以阴道毛滴虫(trichomonas vaginalis,Tv)检测为切入点,构建以量子点和磁微粒技术为基础的夹心法快速检测抗原技术,为同步多项抗原检测作准备。方法针对Tv特异性黏附蛋白AP33制备其抗体,以碳二亚胺交联法,分别连接抗体-量子点及抗体-磁微粒加入待测抗原AP33结合后,在荧光显微镜下观测结果。结果确立了本检测系统的最优条件,对AP33的检测与几种阴道常见病原菌无交叉反应,检测限为0.05μg/ml。结论成功构建了量子点磁微粒检测技术,对Tv抗原检测,特异度为90%,准确率为88%。Objective To establish a new double-antibody sandwich ELISA to detect the antigen AP33 （a 33 kDa adhesive protein） of Trichomonas vaginalis based on the quantum dots and magnetic beads. Methods After BALB/c mice were inmmnized by AP33, the multiclonal antibodies in the antiserum was conjugated with the quantum dots and magnetic beads by carbon diimine crosslinking method respectively. Then the antibodies combined with magnetic beads were coated to microwell of plate as capturing antibody, and the antibodies bound to quantum dots were regarded as the marked antibody which can be directly observed under fluorescence micro- scope and quatitated by spectrofluorometry. The specificity and sensitivity of our established system were investigated. Results AP33 was successfully detected at the concentration as low as 50 ng/ml by this with-filling method. No crossreaction was observed when this system was used to detect Triehomonas vaginalis and other common bacteria in the vagina. The accuracy was 88% and the specificity was 90%. Conclusion This new double-antibody sandwich ELISA to detect Trichomonas vaginalis is successfully prepared and of sound specificity and sensitivity.

关 键 词：量子点 磁微粒 阴道毛滴虫 抗原检测 

分 类 号：R319[医药卫生—基础医学] R382.21