Protein microarray based on dot immunogold filtration assay

    YANG Hao1£¬LI Ding1£¬ HAN Fengª²Chan1£¬LUO Jin1£¬XIA Lin1£¬WANG Xiaoª²Ming2£¬YAN Xiaoª²Jun1

    1PLA Institute of Genetic Diagnosis£¬Fourth Military Medical University£¬Xi¬ðan 710033£¬China£¬ 2Shenzhen Synogene Co. Ltd£¬Shenzhen 518053£¬China

    ¡¾Abstract¡¿ AIM: To develop a cheap and easyª²toª²use protein microarray based on dot immunogold filtration assay for parallel analysis of several targets in serum. METHODS: The human IgG and several antigens were printed on activated nitrocellulose membrane with robotics. After blocked with PBS, the membrane printed was underlaid by macromolecular bibulous material. The patients¡¯ sera samples and immunogold was instilled in the membrane respectively, the relative gray level of human IgG bound to the printed antigens were analysed by CCD reader. The results were compared with ELISA kits. RESULTS: The range of detection for IgG was from 1-50 ng in our microarray. The detection limits was set by the mean of 200 negative serumal samples plus triplicate standard deviation.We assayed 186 random sera, compared with ELISA kit, there was no significance between the two methods, and >90% sensitivity and specificity was obtained. CONCLUSION: Compared with ELISA kits, the microassay performs well equivalently. Furthermore, the microassay is easyª²toª²use and have important advantage in cost and time. It is suitable for clinical diagnosis. ......