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

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.

Rapid detection of avian influenza virus H5N1 in chicken tracheal samples using an impedance aptasensor with gold nanoparticles for signal amplification.

Highly pathogenic avian influenza virus H5N1 is a continuous threat to public health and poultry industry. The recurrence of the H5N1 led us to develop a robust, specific, and rapid detection method for the virus. In this study, an impedance aptasensor was developed for the virus detection using specific H5N1 aptamer and a gold interdigitated microelectrode. Streptavidin was immobilized on the microelectrode surface and biotin labeled H5N1 aptamer was bound to the immobilized streptavidin. The microelectrode was blocked with the polyethylene glycol and the bound aptamer captured the virus. The impedance change caused by the captured virus was measured using an impedance analyzer. To enhance impedance signal, a nanoparticle-based amplifier was designed and implemented by forming a network-like gold nanoparticles/H5N1-aptamer/thiocyanuric acid. The detection limit of the impedance aptasensor was 0.25 HAU for the pure virus and 1 HAU for the tracheal chicken swab samples spiked with the H5N1 virus. The detection time of aptasensor without employing the amplifier was less than an hour. The amplifier increased impedance by a 57-fold for the 1 HAU samples. Only negligible impedance change was observed for non-target viruses such as H5N2, H5N3, H7N2, H1N1, and H2N2. This aptasensor provides a foundation for the development of a portable aptasensor instrument.