A microfluidic biosensor for rapid simultaneous detection of waterborne pathogens.

A microfluidic based biosensor was investigated for rapid and simultaneous detection of Salmonella, Legionella, and Escherichia coli O157:H7 in tap water and wastewater. The biosensor consisted of two sets of focusing electrodes connected in parallel and three sets of interdigitated electrodes (IDE) arrays. The electrodes enabled the biosensor to concentrate and detect bacteria at both low and high concentrations. The focusing region was designed with vertical metal sidewall pairs and multiple tilted thin-film finger pairs to generate positive dielectrophoresis (p-DEP) to force the bacteria moving toward the microchannel centerline. As a result, the bacterial pathogens were highly concentrated when they reached the detection electrode arrays. The detection IDE arrays were coated with three different antibodies against the target bacterial pathogens and a cross-linker to enhance the binding of antibodies to the detection electrode. As the binding of bacterial pathogen to its specific antibodies took place, the impedance value changed. The results demonstrated that the biosensors were capable of detecting Salmonella, Legionella, and E. coli 0157:H7 simultaneously with a detection limit of 3 bacterial cells/ml in 30 - 40 min.

An impedance biosensor for simultaneous detection of low concentration of Salmonella serogroups in poultry and fresh produce samples.

This paper reports the design, fabrication and testing of a microfluidic based impedance biosensor for rapid and simultaneous detection of three Salmonella serogroups. The microfluidic device consists of three microchannels, each one includes a region for focusing the Salmonella cells into the centerline of the microchannel and direct them toward the sensing region to obtain highly concentrated samples using positive dielectrophoresis force. A region for bacteria sensing consists of interdigitated electrode (IDE) array with 10 pairs of fingers. Three types of Salmonella antibodies (type B, D and E) were mixed separately with the cross-linker (Sulfo-LC-SPDP) to enhance the immobalization of the antibodies to the detection electrodes. The electrode surfaces was then functionalized with the three mixtures, one for each channel. As target antigen binds to the antibody, it results in impedance change. The Salmonella samples were spiked with Salmonella type B, introduced into the biosensor via the sample inlet into the focusing region, and then toward the sensing region where they bind to the immobilized antibody, causing a change in the impedance. The performance of the devices was tested using single Salmonella serotype B and two Salmonella serotypes B, and D, with a limit of detection of 7 cells/ml. The biosensor was also able to differentiate live from dead bacteria eliminating the false positive results. Finally, the device was also able to detect Salmonella selectively when other type of pathogen was present.