应用量子点及磁微粒技术快速检测阴道毛滴虫抗原.docVIP

应用量子点及磁微粒技术快速检测阴道毛滴虫抗原 李 蒙，蒲晓允，李 玲，余娟春（第三军医大学新桥医院检验科，重庆 400037） 摘 要:目的 以阴道毛滴虫（richomonas 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，检测为0.05μg/m。结论 量子点磁微粒检测技术对Tv抗原。 关键词量子点 磁微粒 阴道毛滴虫 抗原检测Rapid detection of Trichomonas vaginalis antigens by quantum dots and magnetic beads LI Meng, PU Xiao-yun, LI Ling, YU Juan-chun (Department of Laboratory Medicine, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China) Abstract： 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 immunized by AP33, the multiclonal antibodies in the antiserum was conjungated 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 microscope and quatitated by spectrofluorometry. The specificity and sensitivity of our established system were investigated. Results AP33 was successfully detectede at the consentration as low as 50 ng/ml by this with-filling method. No crossreaction was observed when this system was used to detect Trichomonas 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. Key words: quantum dots; magnetic beads; Trichomonas vaginalis; antigen detection 阴道毛滴虫（richomonas vaginalis,Tv）是一种常见的寄生于人体泌尿生殖系统的鞭毛虫。主要寄生于女性阴道、尿道及男性尿道和前列腺内，引起以性传播为主的泌尿生殖道传染性疾病，如滴虫性阴道炎、尿道炎和前列腺炎。阴道毛滴虫是目前妇科最为常见的引起阴道炎和非淋菌性尿道炎的病原菌之一[1