A Novel Activated Biochar-Based Immunosensor for Rapid Detection of E. coli O157:H7.

Abstract

E. coli O157:H7, one of the major foodborne pathogens, can cause a significant threat to the safety of foods. The aim of this research is to develop an activated biochar-based immunosensor that can rapidly detect E. coli O157:H7 cells without incubation in pure culture. Biochar was developed from corn stalks using proprietary reactors and then activated using steam-activation treatment. The developed activated biochar presented an enhanced surface area of 830.78 m2/g. To develop the biosensor, the gold electrode of the sensor was first coated with activated biochar and then functionalized with streptavidin as a linker and further immobilized with biotin-labeled anti-E. coli polyclonal antibodies (pAbs). The optimum concentration of activated biochar for sensor development was determined to be 20 mg/mL. Binding of anti-E. coli pAbs with E. coli O157:H7 resulted in a significant increase in impedance amplitude from 3.5 to 8.5 kΩ when compared to an only activated biochar-coated electrode. The developed immunosensor was able to detect E. coli O157:H7 cells with a limit of detection of 4 log CFU/mL without incubation. Successful binding of E. coli O157:H7 onto an activated biochar-based immunosensor was observed on the microelectrode surface in scanning electron microscopy (SEM) images.