Highly Sensitive in vitro Biosensor for Enterotoxigenic Escherichia coli Detection Based on ssDNA Anchored on PtNPs-Chitosan Nanocomposite

Abstract

Detection of Enterotoxigenic Escherichia coli in various biological samples has tremendous importance in human health. In this direction, we have designed a label free electrochemical biosensor for highly selective detection of Escherichia coli through detecting ST gene. The ability of sensor probe to detect STG was confirmed using polymerase chain reaction. The biosensor was fabricated based on STG specific probes immobilized on platinum nanoparticles chitosan nanocomposite on screen printed carbon electrode, which was characterized by cyclic voltammetry, transmission electron microscopy, and fourier transform infrared spectroscopy. A highly sensitive label free sensing was achieved by analyzing STG hybridization using electrochemical impedance spectroscopy (EIS) technique. The EIS analysis showed a significant increase in charge transfer resistance after STG interaction with the highly selective ssDNA probe immobilized on the nanocomposite film. The increase in charge transfer resistance was evaluated for varying concentrations of STG, which shows a dynamic range between 1.0×10−12 and 1.0×10−4 with the detection limit of 3.6×10−14 M (RSD<4.5 %). The regeneration of sensor probe was also studied and interference due to non-target sequences was evaluated to ensure the selectivity of the designed sensor. The practical applicability of sensor probe was also analyzed by detecting the STG from the bacteria present in surface water. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim