Rapid Detection of Enterohaemorrhagic E.coli Using Phage-Based Bioluminescent Assay

Abstract

Introduction: There is a need for a reliable, time-saving, and specific detection assay for coliforms, environmental E. coli, and Enterobacteriaceae worldwide. Aim: To innovate a new principle of phage-based rapid diagnostic test for detecting E. coli bacteria in short time and low titer. Materials and Methods: A phage mixture of 200 E. coli specific phages, including 22 specific for Enterohaemorrhagic E.coli (EHEC), were used in a new detection platform, a phage-based Adenylate Kinase Bioluminescence Assay (AKBA). Ten EHEC E. coli and 30 universal E.coli isolates were used for AKBA assay. Results: AKBA showed positive detection of E.coli bacteria at 103 CFU in just 20 minutes. The phage-based detection was highly specific at strain level of E.coli. The sensitivity and specificity of AKBA was 74% and 78%, respectively. Conclusion: A rapid and strain-specific diagnostic test was prepared for E.coli by using coliphages. The significance and impact of the study shows that it might be feasible to formulate a phage-based assay against any Gram negative or positive bacteria using the same approaches of the current AKBA assay with slight modifications.

Keywords

• adenylate kinase
• bacteriophage
• bioluminescence
• rapid diagnostic tests