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

• Published in: Fresenius' journal of analytical chemistry 2001-02, Vol.369 (3-4), p.295-301
• Main Authors: Stokes, D L, Griffin, G D, Vo-Dinh, T
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Biosensing Techniques; Equipment Design; Escherichia coli; Escherichia coli - isolation & purification; Immunoassay - instrumentation; Immunoassay - methods; Milk - microbiology; Reproducibility of Results; Sensitivity and Specificity
• ISSN: 0937-0633; 1432-1130
• DOI: 10.1007/s002160000660

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.