Specific and targeted detection of viable Escherichia coli O157:H7 using a sensitive and reusable impedance biosensor with dose and time response studies

摘要

A gold interdigitated microelectrode (IME) impedance biosensor was fabricated for the detection of viable Escherichia coli O157:H7. This sensor was fabricated using lithography techniques. The surface of the electrode was immobilized with anti-E. coli IgG antibodies. This approach is different from other studies where the change in impedance is measured in terms of growth of bacteria on the electrode, rather then the antibody/antigen bonding. The impedance values were recorded for frequency ranges between 100 Hz and 10 MHz. The working range of the dose response for this device was found to be between 2.5 × 10 ~4 CFU ml ~(-1) and 2.5 × 10 ~7 CFU ml ~(-1). The time response studies indicated that antibody/antigen binding is not a function of time, but can decrease if excess times are allowed for binding. It was observed that the impedance values for 60 min antibody/antigen binding were higher than the impedance values for 120 min binding time. The main advantages of the reported device are that, it provides for both qualitative and quantitative detection in 3 h while other impedance sensors reported earlier may take up to 24 h for detection. If enrichment steps are required then it may take 3-4 days to infer the results. This sensor can be used to detect different types of bacteria by immobilizing the antigen specific antibody. Most of the sensors are not reusable since they either use enzymes or enrichment steps for detection but this device can be reused, following a cleaning protocol which is easy to follow. Each device was used at least five times. The simplicity of this sensor and the ease of fabrication make this sensor a useful alternate to the microfluidics and enzyme based impedance sensors, which are relatively more difficult to fabricate, need programmable fluidic injection pumps to push the sample through the channel, suffer from limitation of coagulation and are difficult to clean.