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Recombinant Phage Tail Fiber Protein Gp47 Enables Broad-Spectrum Fluorescent Detection of Pseudomonas aeruginosa
Honglin Yang1, Qinchen Liao1, Kuixun Chen1
1Key Laboratory of Luminescence Analysis and Molecular Sensing (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
Abstract:
Pseudomonas aeruginosa (P. aeruginosa) has long been one of the leading opportunistic pathogens responsible for serious healthcare-associated infective diseases. Development of rapid and accurate detection techniques for this pathogen is highly desired to achieve efficient prevention and control. This study developed a fluorescent detection technique targeting this pathogen by using recombinant phage tail fiber protein Gp47 as the recognition element. This protein was identified from a phage PAB1 isolated from hospital wastewater through genomic sequencing and bioinformatic analysis. It was then produced in its functional form via recombinant expression and purification. Gp47 exhibited broad-spectrum binding capability toward P. aeruginosa strains, recognizing all 30 clinical isolates tested, while completely lacking lytic activity. Thus, it effectively overcame the key limitations of the whole phage, including the narrow host range and tendency to lyse target bacteria. A sandwich method was constructed to detect P. aeruginosa by using Gp47 conjugated magnetic beads as capture carriers and fluorescein isothiocyanate labeled antimicrobial peptide as signal reporter. The method responded to P. aeruginosa linearly from 3.2 × 102 to 3.2 × 106 CFU/mL, with a limit of detection of 109 CFU/mL and ideal specificity. It has been employed for detecting P. aeruginosa in complex matrices such as physiological saline, human urine, and milk powder with acceptable recoveries ranging from 71.23% to 110.87%. This work provides a high-performance recognition element for P. aeruginosa and helps develop valuable diagnostic tools for infective diseases.
Insights
A new fluorescent detection method uses recombinant phage protein Gp47 to rapidly identify Pseudomonas aeruginosa (P. aeruginosa). This technique offers high specificity and sensitivity for detecting this opportunistic pathogen in various samples.
Area of Science:
- Microbiology
- Biotechnology
- Diagnostic Development
Background:
- Pseudomonas aeruginosa (P. aeruginosa) is a major opportunistic pathogen causing healthcare-associated infections.
- Rapid and accurate detection methods are crucial for controlling P. aeruginosa infections.
Purpose of the Study:
- To develop a novel fluorescent detection technique for P. aeruginosa.
- To utilize recombinant phage tail fiber protein Gp47 as a specific recognition element.
Main Methods:
- Identified and recombinantly expressed phage protein Gp47 from phage PAB1.
- Developed a sandwich assay using Gp47-conjugated magnetic beads and a fluorescently labeled antimicrobial peptide.
- Tested detection in complex matrices like saline, urine, and milk powder.
Main Results:
- Gp47 demonstrated broad-spectrum binding to P. aeruginosa clinical isolates without lytic activity.
- The assay detected P. aeruginosa linearly from 3.2 × 10^2 to 3.2 × 10^6 CFU/mL with a limit of detection of 109 CFU/mL.
- Acceptable recovery rates (71.23%–110.87%) were achieved in complex matrices.
Conclusions:
- Recombinant Gp47 is a high-performance recognition element for P. aeruginosa detection.
- The developed fluorescent assay provides a sensitive, specific, and versatile tool for diagnosing P. aeruginosa infections.
