结直肠癌血清肿瘤标记物液相芯片制备条件的优化及在CEA抗原检测中的初步应用  

Optimization of a Liquid Chip System for the Detection of Serum Biomarkers of Colorectal Cancer and Its Application

作　　者：张华宁[1] 高雪芹[1] 韩金祥[1] 黄海南[1] 

机构地区：[1]山东省医药生物技术研究中心卫生部生物技术药物重点实验室山东省现代医用药物与技术重点实验室,济南250062

出　　处：《中国生物工程杂志》2006年第12期81-85,共5页China Biotechnology

基　　金：山东省自然科学基金资助项目(Y2003C42)

摘　　要：液相芯片技术由于其高通量,灵敏度高,信噪比高,液相条件下反应,操作简便,耗时短等优点,已被美国FDA批准成为临床的检测手段。主要介绍了结直肠癌血清肿瘤标记物液相芯片制备条件的优化及其在CEA抗原检测中的初步应用。首先将CEA抗原的捕获抗体与微球载体进行偶联,制备液相芯片,然后对影响反应的微球与抗原的反应时间,生物素化检测抗体的浓度及avidin-PE荧光染料的反应浓度等因素进行正交设计,确定出最优的反应条件;用该液相芯片反应体系检测55例临床样本,与ELISA试剂盒检测结果相比:在同样的样本浓度范围内,两者的检测结果基本一致,但液相芯片检测的浓度范围更大而且液相芯片可将多种肿瘤标记物在一个反应进行检测,节省检测的时间和人力。Liquid chip technology have been licensed to be used in clinic because of its advantage of high-throughput, high-sensitivity, good signal to noise ratio, reaction in liquid phase, convenient operation and short time consuming, etc. The optimization of a liquid chip system for the detection of serum biomarkers of colorectal tumour and initial application in the detection of CEA were studied. The optimized reaction conditions of liquid chip were determined through orthogonal design after it was prepared. The results showed that the consuming reaction time of the coated antibody and the antigen was 1 hour. The microspheres, biotinylated detecion antibody and the consuming complexes and avidin-PE time of the microspheres and the biotinylated tested antibody was lhour, lhour and 15minutes respectively, the consuming time of the complexes and avidin-PE was fifteen minutes, The optimized dilution of the biotinylated tested detection antibody was 1 ： 300 and the optimized concentration of avidin-PE was 12μg/ml. Totally 55 clinical samples were detected by the liquid chip and by Enzyme-Linked Immunosorbent Assay （ELISA） simultaneously and the results of the two methods were compared. The results of the two methods showed good correlation between positive and negative samples but the detection limits and the dynamic ranges of the liquid chip method were more sensitive and -ruder than those of the ELISA. The multiple tumour biomarkers may be detected simultaneously and the time of clinical test and manpower requirements were reduced by the liquid chip method.

关 键 词：肿瘤标记物 液相蛋白芯片 癌胚抗原 酶联免疫吸附实验 

分 类 号：R730.4[医药卫生—肿瘤]