Simultaneous detection of E. coli K12 and S. aureus Using a Continuous Flow Multijunction Biosensor

Rapid detection and identification of potentially harmful bacteria is ideal for food manufacturers to prevent foodborne illness outbreaks. Continuous monitoring method of foodborne pathogens levels and trends in food gives real-time results. Therefore, the objectives of this study were to fabricate and characterize the continuous flow multijunction biosensor for simultaneous detection of Escherichia coli K12 and Staphylococcus aureus. Junction biosensors were fabricated using gold plated tungsten wires coated with polyethylenimine and single walled carbon nanotubes. Each junction was functionalized with streptavidin and biotinylated antibodies specific to E. coli K12 and S. aureus. Then, single or 2 biosensors for each targeted analyte were connected to tubing, perpendicular to the flow direction. Pure serial diluted samples of E. coli K12 and S. aureus and microbial cocktail samples were continuously pumped at a 0.0167 mL/s into the detection zone. Changes in the electric current by biorecognition reactions between antibody and antigens were calculated. The developed junction sensor coupled with the fluidic channel showed the enhancement of the electric signal responses for detection of E. coli K12, compared to the stationary sensor. A linear regression was observed for both the E. coli and S. aureus functionalized array sensors in the detection range of 102 to 105 CFU/mL. Multiplexed detection of bacteria at the sensing levels as low as 102 CFU/mL for E. coli K12 and S. aureus was achieved within 2 min. Therefore, the continuous flow multijunction biosensor shows potential for rapid and continuous multiplexed detection of foodborne pathogens.