Development of a multi-channel immunosensor for determnination of serum hepatic fibrosis markers

Honghu Huang, Jia Zhou, Yipin Huang, M. Bao
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Abstract

In this study, basing on electrochemistry and MEMS technology we develop a novel biosensor with multi channels to detect three markers (hyaluronic acid (HA), Laminin (LN), type IV collagen (IV-C) ) so as to evaluate the liver disease. With MEMS technology, a chip including a CE, a RE and 6 gold electrodes is fabricated for detection of at most 6 markers simultaneously. Different antibodies of serum markers are embedded in poly(o-phenylenediamine) to form semi-insulated membranes on the gold electrodes, separately by electrochemical polymerization. It is grounded on a mechanism of detecting the variation of the peak current caused by the immunoreactions between the markers and their corresponding antibodies. Concentrations of HA, LN and IV-C in 140 samples are tested both by the sensor and RIA, namely, sensor group and RIA group. Compared the two groups of results, it is found that the two measures have similar sensitivities and consistency while the sensor has significantly reduced the response time as well as the cost. The coefficients of variation of the sensor are less than 20% for both inter and intra-assay with error less than 20%, while for RIA, coefficients of variation for intra-assay less than 10% and inter-assay less than 15% with error less than 20%
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多通道免疫传感器测定血清肝纤维化标志物的研制
本研究基于电化学和MEMS技术,开发了一种新型的多通道生物传感器,检测三种标志物(透明质酸(HA),层粘连蛋白(LN), IV型胶原蛋白(IV- c)),以评估肝脏疾病。采用MEMS技术,制作了一个芯片,包括CE, RE和6个金电极,最多可同时检测6个标记。将不同的血清标记物抗体分别包埋在聚邻苯二胺中,通过电化学聚合在金电极上形成半绝缘膜。它基于一种检测标记物与其相应抗体之间的免疫反应引起的峰值电流变化的机制。140个样品中HA、LN和IV-C的浓度分别通过传感器和RIA检测,即传感器组和RIA组。对比两组测量结果,发现两种测量方法具有相似的灵敏度和一致性,同时传感器显著降低了响应时间和成本。该传感器的组间和组内变异系数小于20%,误差小于20%,而RIA组内变异系数小于10%,组间变异系数小于15%,误差小于20%
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