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使用基于微流控的抗体生物芯片检测系统检测大肠杆菌。

Detection of E. coli using a microfluidics-based antibody biochip detection system.

作者信息

Stokes D L, Griffin G D, Vo-Dinh T

机构信息

Advanced Monitoring Development Group, Life Sciences Division, Oak Ridge National Laboratory, TN 37831-6101, USA.

出版信息

Fresenius J Anal Chem. 2001 Feb;369(3-4):295-301. doi: 10.1007/s002160000660.

Abstract

This work demonstrates the detection of E. coli using a 2-dimensional photosensor array biochip which is efficiently equipped with a microfluidics sample/reagent delivery system for on-chip monitoring of bioassays. The biochip features a 4 x 4 array of independently operating photodiodes that are integrated along with amplifiers, discriminators and logic circuitry on a single platform. The microfluidics system includes a single 0.4 mL reaction chamber which houses a sampling platform that selectively captures detection probes from a sample through the use of immobilized bioreceptors. The independently operating photodiodes allow simultaneous monitoring of multiple samples. In this study the sampling platform is a cellulosic membrane that is exposed to E. coli organisms and subsequently analyzed using a sandwich immunoassay involving a Cy5-labeled antibody probe. The combined effectiveness of the integrated circuit (IC) biochip and the immunoassay is evaluated for assays performed both by conventional laboratory means followed by detection with the IC biochip, and through the use of the microfluidics system for on-chip detection. Highlights of the studies show that the biochip has a linear dynamic range of three orders of magnitude observed for conventional assays, and can detect 20 E. coli organisms. Selective detection of E. coli in a complex medium, milk diluent, is also reported for both off-chip and on-chip assays.

摘要

这项工作展示了使用二维光电传感器阵列生物芯片对大肠杆菌的检测,该生物芯片高效配备了微流控样品/试剂输送系统,用于生物测定的芯片上监测。该生物芯片具有一个4×4的独立运行光电二极管阵列,这些光电二极管与放大器、鉴别器和逻辑电路集成在一个单一平台上。微流控系统包括一个0.4 mL的单一反应腔,该反应腔容纳一个采样平台,该平台通过使用固定化生物受体从样品中选择性捕获检测探针。独立运行的光电二极管允许同时监测多个样品。在本研究中,采样平台是一张纤维素膜,将其暴露于大肠杆菌生物体,随后使用涉及Cy5标记抗体探针的夹心免疫测定法进行分析。评估了集成电路(IC)生物芯片和免疫测定法的综合有效性,分别通过传统实验室方法进行测定,然后用IC生物芯片进行检测,以及通过使用微流控系统进行芯片上检测。研究亮点表明,对于传统测定,该生物芯片的线性动态范围为三个数量级,并且能够检测到20个大肠杆菌生物体。还报告了在复杂介质牛奶稀释液中对大肠杆菌进行芯片外和芯片上测定的选择性检测。

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