Abstract: In this study, we designed primers and probes targeting the gyrB gene of Pseudomonas fluorescens, prepared standard plasmids, drew standard curves, and established a real-time polymerase chain reaction (PCR) system for the detection of P. fluorescens. Our results confirmed that the PCR system was specific to P. fluorescens without detection of other tested bacteria. The sensitivity was 14.3 fg/μL and 3.0 × 102 CFU/mL for pure DNA and pure culture, respectively, and was not affected by the background interference at low concentrations. In artificially contaminated samples, P. fluorescens could be detected by appropriate enrichment. The TaqMan-based real-time fluorescence quantitative PCR method proved to be highly specific, sensitive and resistant to interferences.

Key words: Pseudomonas fluorescens; real-time polymerase chain reaction; gyrB gene