Engineered bacteriophage biosensors for rapid detection of pathogenic bacteria

Overview

This research proposes a novel biosensing platform that leverages engineered bacteriophages to detect specific pathogenic bacteria. Bacteriophages are viruses that naturally infect and replicate within bacterial hosts, with different phage strains targeting different bacterial species. By selecting a specific bacteriophage for each target pathogen and engineering it to carry a reporter gene, the platform produces a quantifiable signal only when the phage encounters its intended host. Because phages replicate only within living bacteria, this approach detects viable cells rather than non-living remnants, offering a meaningful advantage over many conventional detection methods.

The platform targets four clinically relevant pathogens: Ralstonia pickettii, Brevundimonas diminuta, Pseudomonas aeruginosa, and Mycobacterium abscessus. These organisms are associated with healthcare-associated infections, contamination of pharmaceutical products and medical devices, and infections in immunocompromised patients. The technology has potential applications in clinical diagnostics, pharmaceutical manufacturing quality control, water and food safety testing, and environmental monitoring.

Technical specifications

Core approach:

• Identification of a specific bacteriophage for each target pathogenic species
• Molecular engineering of each phage to incorporate a reporter gene
• Reporter expression occurs only during active infection of the correct host bacterium
• Signal amplification through natural phage replication within viable host cells

Key features:

• Species-specific detection through natural phage-host specificity
• Detection of viable cells only, reducing false positives from dead bacteria
• Potential for high sensitivity based on demonstrated phage-based detection of E. coli at approximately 10 CFU/ml
• Modular design allowing expansion to additional pathogens by swapping in new phage-reporter pairs

The detection method builds on demonstrated phage engineering techniques from the scientific literature and the investigator's prior experience developing detection methods for Clostridium botulinum and studying phage receptor-based detection systems.

Technology readiness level

This technology is at an early research stage. The proposal outlines a one-year development plan beginning with phage identification for each target pathogen, followed by molecular engineering to introduce reporter genes, and concluding with evaluation of detection capabilities. The approach is grounded in existing literature on reporter-gene phage engineering and the investigator's relevant prior work in pathogen detection. The platform has not yet been validated for the four target organisms, and significant development work remains before commercial or clinical deployment.