Bacteriophage long tail fibre proteins as a biorecognition element in electrochemical biosensors for the detection of Salmonella in lake water.

We developed an electrochemical impedance spectroscopy (EIS) biosensor based on bacteriophage long tail fibre proteins (LTFPs) to capture and detect Salmonella. First, based on the genetic information of a Salmonella enterica bacteriophage (SEP37), we found that the genes ORF99, ORF100, and ORF256 encode proteins associated with host recognition. We used the pETdue1 plasmid to construct ORF99–ORF100 and ORF256 co-expression vectors to express (LTFPs) in Escherichia coli and used purified LTFPs as specific biorecognition elements. Then, we fixed gold nanoparticles (AuNPs), cysteamine (Cys), and LTFPs on a gold disk electrode surface via layer-by-layer assembly to construct an EIS biosensor. Salmonella in the sample can be captured by LTFPs on the electrode surface and reduce the electron transfer sites. We applied the EIS biosensor to detect Salmonella spiked in lake water. It showed a good linear relationship within the concentration range of 10 ∼ 1 × 106 colony-forming units (CFU)/mL. The detection limit was 9 CFU/mL. This method based on bacteriophage LTFPs does not require complex pretreatment steps and extends the application of tail fibre proteins as a biorecognition element in the detection field. [Display omitted] • ORF99-100 and ORF256 were inserted into plasmid to construct co-expression vector. • The long tail fibre protein of phage SEP37 was co-expressed in E. coli. • This protein was used as bio-recognition element to construct an EIS biosensor. • This biosensor can quickly detect Salmonella in lake water samples. • This method allowed a detection limit of 9 CFU/mL for Salmonella. [ABSTRACT FROM AUTHOR]