基于斑点免疫金渗滤法的蛋白微矩阵
斑点免疫金渗滤法,,斑点免疫金渗滤法;蛋白质类;微矩阵,0引言,1材料和方法,2结果,3讨论
Protein microarray based on dot immunogold filtration assayYANG Hao1,LI Ding1, HAN FengChan1,LUO Jin1,XIA Lin1,WANG XiaoMing2,YAN XiaoJun1
1PLA Institute of Genetic Diagnosis,Fourth Military Medical University,Xian 710033,China, 2Shenzhen Synogene Co. Ltd,Shenzhen 518053,China
【Abstract】 AIM: To develop a cheap and easytouse 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 easytouse and have important advantage in cost and time. It is suitable for clinical diagnosis. ......
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