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.

Global Epidemiology of Campylobacter Infection

“…Furthermore, a number of real-time assays are also available for the detection of Campylobacter species, some of which are capable of detecting more than one species at a time, including C. jejuni, C. coli, and C. lari (57). Interestingly, Javed and colleagues also described an assay to detect C. jejuni and C. coli based on the ability of recombinant receptor binding proteins from the C. jejuni bacteriophage NCTC12673 to agglutinate in the presence of these species (276).…”

Modified Bacteriophage S16 Long Tail Fiber Proteins for Rapid and Specific Immobilization and Detection of Salmonella Cells

“…Campylobacter phage RBPs have also been developed as probes for detecting the organism via surface plasmon resonance (SPR) biosensors (41), as well as for a cell agglutination assay (42).…”

Yersinia enterocolitica-Specific Infection by Bacteriophages TG1 and ϕR1-RT Is Dependent on Temperature-Regulated Expression of the Phage Host Receptor OmpF

“…Confocal laser immunofluorescence microscopy was used to visualize the binding of the phage TG1 LTF protein Gp37 to Y. enterocolitica by following methodology described by others (30). Yersinia strains grown in Trypticase soy broth (TSB) at 25°C or 37°C for 24 h were resuspended in wash buffer (50 mM Tris-HCl, pH 7.5), and 10-l volumes were spotted onto clean glass slides.…”