Charge-Directed Immobilization of Bacteriophage on Nanostructured Electrode for Whole-Cell Electrochemical Biosensors

A new type of carbon nanotube (CNT)-based impedimetric biosensing method has been developed for rapid and selective detection of live bacterial cells. A proof-of-concept study was conducted using T2 bacteriophage-based biosensors for electrochemical detection of Escherichia coli B. The T2 bacteriophage (virus) served as the biorecognition element, which was immobilized on polyethylenimine (PEI)-functionalized carbon nanotube transducer on glassy carbon electrode. Charge-directed, orientated immobilization of bacteriophage particles on carbon nanotubes was achieved through covalent linkage of phage capsid onto the carbon nanotubes. The presence of the immobilized phage on carbon nanotube-modified electrode was confirmed by fluorescence microscopy. Electrochemical impedance spectroscopy (EIS) was used to monitor the changes in the interfacial impedance due to the binding of E. coli B to T2 phage on the CNT-modified electrode. The detection was highly selective toward the B strain of E. coli as no signal was observed for the nonhost K strain of E. coli. The present achievable detection limit of the biosensor is 10³ CFU/mL.