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An impedance-based DNA multiplexed biosensor was designed to simultaneously detect Escherichia coli (yaiO gene) and its virulent f17 variant. The thiolated DNA dual probe was self-assembled onto the surface of the gold nanoparticle-modified screen-printed carbon electrode (AuNPs/SPCE) to recognize selected sequences from yaiO and f17 genes. The optimal conditions to prepare the bioelectrode were determined based on the monitoring of the impedimetric response fitted to an equivalent electrical circuit model. The charge transfer resistance of the bioelectrode increased by recognizing the target DNA sequences. The limit of detection was 0.8 fM and 1.0 fM for yaiO and f17 target DNA, respectively, and the linearity ranged from 1 × 10-15 to 1 × 10-7 M with a linear regression coefficient R ≥ 0.995. The nanodevice provided a novel strategy for simultaneous detection of E. coli and its virulence f17 gene with excellent discrimination with a single-base mismatch, two-base mismatch, and non-complementary sequences. Moreover, genomic DNA extracted from E. coli bacteria isolated from diarrheic camel calves and control animals in Tunisia was successfully detected using the as-prepared biosensor with minimal treatment of the extracted DNA samples.Graphical abstract.
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10.1007/s00604-020-04614-y
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Impedimetric DNA E-biosensor for multiplexed sensing of Escherichia coli and its virulent f17 strains.
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