同时检测血清中多种抗体的蛋白质微阵列研究

建立一种用蛋白质微阵列法可同时检测血清中艾滋病毒（HIV）, 丙型肝炎病毒（HCV）和梅毒螺旋体（TP）抗体的方法.将基因工程HIV、HCV和TP 3种融合抗原共价结合于固相载体玻片上，制成蛋白质微阵列，血清样本经稀释、加样、孵育、洗涤后，加上Cy3荧光标记二抗，洗涤，激光共聚扫描，将获得的图片用Bio-discover公司的Imagene专用分析软件进行分析，所获数据在经过自行编写的软件,根据Cutoff值自动生成判断结果.用此蛋白质微阵列系统检测了400例阴性血清，确定了Cutoff值，检测中国药品生物制品检定研究所的HIV, HCV和TP 3种参比品，并与3种ELISA试剂盒的结果进行了比较.蛋白质微阵列的艾滋病毒阳性和阴性符合率均为100%(20/20); 丙型肝炎病毒阳性和阴性符合率均为95%(38/40).梅毒螺旋体参比品阳性符合率为100%(10/10)，阴性符合率为100%(20/20)，蛋白质微阵列与三种ELISA试剂检测国家HIV、TP、HCV参比品（共150份血清样本），结果具有高度的符合率.

Studies on Simultaneously Detecting Multiple Antibodies in The Serum Using Microarray

An optimized method has been established to simultaneously detect multiple antibodies of HIV, TP and HCV in the serum by the way of microarray. Chimeric antigens of HIV, TP and HCV are covalently bound to the surface of glass slide; serum samples are diluted and added to the well of microarray, unbound antibodies are washed away after incubation, and Cy3 labeled second antibody is added to the well followed by incubation and washing. The microarray is scanned by laser confocal scanner, and the resulting images are treated by the software of Imagene (produced by Bio-discover Co.). The data obtained is then treated by the software produced by own; the evaluated result of each sample is finally gained automatically by the Cutoff value of each antigen. 400 normal serum sample are detected and Cutoff value of each antigen are determined. National panels of HIV, TP and HCV had also been detected by this system and the results were compared with those detected by ELISA method. The coordinate ratios of positive and negative samples of HIV panel are both 100%(20/20); these kinds of the coordinate ratios of positive and negative samples of HCV are both 95%(38/40) and the coordinate ratios of positive and negative samples of TP are 100%(10/10) and 100%(20/20) respectively. This result is highly in accord with that of ELISA method.