Objective To establish a real-time recombinase-aid amplification(RAA) method for rapid detection of Pseudomonas aeruginosa. Methods Specific primers and exo probes based on ecfX gene of P.aeruginosa were designed in this study, and the validity of the method was evaluated by sensitivity, specificity and suspected strains detection. Results Real-time RAA was performed successfully at 39 ℃ for 20 min. Only the P.aeruginosa strains but not other bacteria were amplified, showing the good specificity. The limit of detection was 3.0×10³fg genomic DNA per reaction, and 1.0×10³ CFU P.aeruginosa pure culture per reaction. The developed real-time RAA was further evaluated on 36 suspected of P.aeruginosa, which were identified successfully to be P.aeruginosa. The detection result were the same with those of a real-time PCR assay and the VITEK 2 Compact. Conclusion The developed real-time RAA assay is a rapid, simple and reliable tool for accurate detection of P.aeruginosa of diverse origins.