# Rapid On-Site Detection of Pseudomonas aeruginosa via ecfX-Targeted Loop-Mediated Isothermal Amplification

## TL;DR

A new rapid test detects Pseudomonas aeruginosa using a specific gene target, offering quick and accurate results for clinical and environmental use.

## Contribution

A novel fluorescence-based LAMP method targeting the ecfX gene enables rapid and specific detection of Pseudomonas aeruginosa.

## Key findings

- The optimal primer set EC2 achieved high sensitivity and specificity for Pseudomonas aeruginosa detection.
- The LAMP assay showed no cross-reactivity with non-PA pathogens and comparable detection limits to PCR.
- The method was validated with bacterial strains and environmental samples, confirming its reliability and feasibility.

## Abstract

Pseudomonas aeruginosa (PA) is a significant pathogen of clinical concern that is frequently associated with multidrug resistance, leading to respiratory tract, wound, and hospital-acquired infections. To enable rapid and accurate detection, we developed a fluorescence-based loop-mediated isothermal amplification (LAMP) method, targeting the PA-specific ecfX gene. Among ten primer sets designed, the optimal set (EC2) was identified, and reaction conditions were optimized (Bst polymerase 320 U/mL, Mg2+ 8 mM, dNTP 1.4 mM, inner/outer primer ratio 1:8, 64 °C, 20 min). The assay demonstrated a detection limit that was comparable to a real-time polymerase chain reaction and immunochromatographic assays, but with a markedly reduced turnaround time. No cross-reactivity was observed with non-PA pathogens, and reproducibility tests confirmed high stability. In addition, the reliability of the results was further verified using 60 standard bacterial strains, and the feasibility of the assay was validated with 2 real soil samples and 1 water sample. This LAMP method offers a simple, rapid, and sensitive tool for on-site detection of PA, with potential applications in clinical diagnostics and public health surveillance.