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

中国生物工程杂志

技术与方法

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

张华宁高雪芹韩金祥黄海南

山东省医药生物技术研究中心山东省医药生物技术研究中心山东省医学科学院生物技术研究中心信阳师范学院生命科学学院

Optimization of a liquid chip system for serum biomarkers of colorectal tumour and initial application in the detection of CEA

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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。本文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。本研究首先将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。

Abstract: 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 and so on. This study focused on the optimization of a liquid chip system for the detection of serum biomarkers of colorectal tumour and initial application in the detection of CEA. The optimized reaction conditions of liquid chip were determined through orthogonal design . The results showed that the reaction time of the coated antibody and the antigen,、the microspheres and the biotinylated detecion antibody and the complexes and avidin-PE was 1 hour, 1hour and 15minutes respectively.The optimized dilution of the biotinylated 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 in both positive and negative samples but the detection limits and the dynamic ranges of the liquid chip method were more sensitive and wider 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．

收稿日期: 2006-08-21 出版日期: 2006-12-25

通讯作者: 张华宁

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引用本文:

张华宁,高雪芹,韩金祥,黄海南. 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[J]. 中国生物工程杂志, .

. Optimization of a liquid chip system for serum biomarkers of colorectal tumour and initial application in the detection of CEA. China Biotechnology, .

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https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I12/81

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