Determination of carcinoembryonic antigen in human sera by integrated bead bed immunoasay in a microchip for cancer diagnosis
Determination of carcinoembryonic antigen in human sera by integrated bead bed immunoasay in a microchip for cancer diagnosis
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DOI：
10.1021/ac000991z
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发表时间：
2001-03-15
影响因子：
7.4
通讯作者：
Kitamori, T
中科院分区：
文献类型：
--
作者：
Sato, K;Tokeshi, M;Kitamori, T
在微芯片上构建珠床免疫分析系统，并应用于测定癌胚抗原（CEA），这是一种常用的结肠癌标志物。将预涂有抗癌胚抗原抗体的聚苯乙烯珠引入微通道，然后将含有癌胚抗原的血清样品、第一抗体和与胶体金偶联的第二抗体依次反应，得到固定在珠表面的抗原抗体复合物，使用热透镜显微镜（TLM）进行检测。一个高度选择性和灵敏度的测定超微量的CEA在人血清中的夹心免疫测定系统，需要三个抗体的测定。检测限比常规ELISA低几十倍。用该系统检测13例患者血清，与常规ELISA法的相关系数为0.917。集成减少了抗原抗体反应所需的时间约为1%，从而缩短了总的分析时间从45小时到35分钟。此外，麻烦的操作所需的常规异质性免疫分析可以大大简化。这种基于微芯片的诊断系统是第一个基于微芯片的系统，它实际上可用于临床诊断，分析时间短，灵敏度高，程序简单。
A bead-bed immunoassay system was structured on a microchip and applied to determine carcinoembryonic antigen (CEA), which is a commonly used marker of colon cancer. Polystyrene beads precoated with anti-CEA antibody were introduced into a microchannel, and then a serum sample containing CEA, the first antibody, and the second antibody conjugated with colloidal gold were reacted successively, The resulting antigen-antibodies complex, fixed on the bead surface, was detected using a thermal lens microscope (TLM). A highly elective and sensitive determination of an ultratrace amount of CEA in human sera was made possible by a sandwich immunoassay system that needs three antibodies for an assay. A detection limit dozens of times lower than the conventional ELISA was achieved. Moreover, when serum samples for 13 patients were assayed with this system, there was a high correlation (r = 0.917) with the conventional ELISA. The integration reduced the time necessary for the antigen-antibody reaction to similar to1%, thus shortening the overall analysis time from 45 h to 35 min. Moreover, troublesome operations required for conventional heterogeneous immunoassays could be much simplified. This microchip-based diagnosis system is the first microchip-based system that is practically useful for clinical diagnoses with short analysis time, high sensitivity, and easy procedures.