液相蛋白质芯片检测血清C反应蛋白方法的建立

目的 建立液相蛋白质芯片检测血清C反应蛋白（CRP）的方法，探讨该方法诊断冠心痛的临床应用价值。方法 用聚苯乙烯荧光微珠，将抗CRP单克隆抗体包被在微珠上，将另一针对CRP不同抗原表位的单克隆抗体生物素化；包被好的微珠加入到96孔板中，将待测血清分别加入各孔，用CRP标准品建立标准曲线，各孔中加入生物素化抗CRP单抗和PE荧光素标记的链霉亲合素。在Bio-Plex液相蛋白质芯片分析仪上检测296份血清样品。同一血清标本同时用免疫比浊法测定。比较两种方法对CRP检测的敏感性和特异性。结果 液相蛋白质芯片检测CRP方法对诊断冠心病的敏感性（78，3％）和特异性（87．1％）较免疫比浊法的敏感性（63．0％）和特异性（86．5％）高，χ^2=28，94，P〈0．001。结论 建立了一种检测血清CRP的新方法，对诊断冠心痛有临床应用价值。

Liquid chip:a new method for detection of serum high-sensitivity C-reactive protein

Objective To establish a new method using liquid chip for detection of serum high-sensitivity C-reactive protein (CRP) and estimate its values in clinical application for diagnosis of coronary heart disease (CHD).Methods A monoclonal antibody to CRP was coupled to the fluorescent beads of carboxylated polystyren and another monoclonal antibody to different epitope of CRP was biotinylated.The antibody-coupled beads were added into each well of 96-well microtiter plate and the diluted standard or sample was in turn added to the corresponding wells.Following keeping the plate in dark streptavidin-PE were added into each well.The plate was then proceeded on Bio-Plex protein array system to read the results.A total of 296 samples were detected by this method and immunoturbidimetry simultaneously.The sensitivity and specificity of the 2 methods were compared.Results The sensitivity of the established liquid chip method was 78.3% and the specificity was 87.1%,while in the immunoturbidimetry were 63.0% and 86.5% respectively.Conclusion Using the liquid chip system to detect serum CRP is a credible method.The establishment of this method will offer feasibility for using liquid chip system to detect multiple protein markers in diagnosis of CHD and other diseases.