Dual-site recognition of Pseudomonas aeruginosa using polymyxin B and bacteriophage tail fiber protein

Abstract

As one of the top three opportunistic pathogens, Pseudomonas aeruginosa (P. aeruginosa) has long accounted for hospital-acquired infections with high risk of death. In this work, a fluorescent method based on a dual-site recognition mode was developed for rapid assay of P. aeruginosa. Employing its strong binding capability towards lipid A on the outer membrane of Gram-negative bacteria, polymyxin B acted as one recognition element for P. aeruginosa. To overcome the poor binding specificity of polymyxin B, a recombinant bacteriophage tail fiber protein was expressed and employed as a species-specific recognition element for the target pathogen. Thus a dual-site recognition mode was developed for specific assay of P. aeruginosa species by using fluorescein isothiocyanate as a fluorescent probe. The target pathogen can be assayed within a broad dynamic range from 2.0 × 103 CFU mL−1 to 2.0 × 107 CFU mL−1. Due to the ideal specificity of tail fiber protein, the method is capable of excluding the interference from other Gram-negative bacteria and all Gram-positive bacteria. It has been employed for assaying P. aeruginosa in various types of sample matrixes inclusive of lake water, physiological saline injection, human urine and milk. The acceptable assay results demonstrate its promising prospect for practical application in various areas such as environmental hygiene, medical diagnosis, as well as drug and food safety.