title: Bacteriophage receptor binding protein based assays for the simultaneous detection of Campylobacter jejuni and Campylobacter coli.
abstract: Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of foodborne gastroenteritis which is occasionally followed by a debilitating neuropathy known as Guillain-Barré syndrome. Rapid and specific detection of these pathogens is very important for effective control and quick treatment of infection. Most of the diagnostics available for these organisms are time consuming and require technical expertise with expensive instruments and reagents to perform. Bacteriophages bind to their host specifically through their receptor binding proteins (RBPs), which can be exploited for pathogen detection. We recently sequenced the genome of C. jejuni phage NCTC12673 and identified its putative host receptor binding protein, Gp047. In the current study, we localized the receptor binding domain to the C-terminal quarter of Gp047. CC-Gp047 could be produced recombinantly and was capable of agglutinating both C. jejuni and C. coli cells unlike the host range of the parent phage which is limited to a subset of C. jejuni isolates. The agglutination procedure could be performed within minutes on a glass slide at room temperature and was not hindered by the presence of buffers or nutrient media. This agglutination assay showed 100% specificity and the sensitivity was 95% for C. jejuni (n = 40) and 90% for C. coli (n = 19). CC-Gp047 was also expressed as a fusion with enhanced green fluorescent protein (EGFP). Chimeric EGFP_CC-Gp047 was able to specifically label C. jejuni and C. coli cells in mixed cultures allowing for the detection of these pathogens by fluorescent microscopy. This study describes a simple and rapid method for the detection of C. jejuni and C. coli using engineered phage RBPs and offers a promising new diagnostics platform for healthcare and surveillance laboratories.
authors.affiliation: Alberta Glycomics Centre and Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada. majaved@ualberta.ca
authors.forename: Muhammad A
authors.initials: MA
authors.lastname: Javed
authors.forename: Somayyeh
authors.initials: S
authors.lastname: Poshtiban
authors.forename: Denis
authors.initials: D
authors.lastname: Arutyunov
authors.forename: Stephane
authors.initials: S
authors.lastname: Evoy
authors.forename: Christine M
authors.initials: CM
authors.lastname: Szymanski
mesh_terms: D000372:Agglutination Tests / Q000379:methods*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism*; D017000:Campylobacter coli / Q000302:isolation & purification* / Q000648:ultrastructure / Q000821:virology; D016123:Campylobacter jejuni / Q000302:isolation & purification* / Q000648:ultrastructure / Q000821:virology; D017931:DNA Primers / Q000235:genetics; D005759:Gastroenteritis / Q000382:microbiology*; D049452:Green Fluorescent Proteins; D006801:Humans; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012680:Sensitivity and Specificity
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins