Detection of Pseudomonas anguilliseptica from Olive Flounder Paralichthys olivaceus Using Real-time PCR with a TaqMan Fluorescent Probe

抄録

Pseudomonas anguilliseptica is a dreadful fish pathogen causing severe problem in olive flounder Paralichthys olivaceus farms in Jeju Island, Korea. Diagnosis of this disease required a time-consuming and labor intensive culturing step. Hence more rapid and sensitive detection methods are needed. In this study, a TaqMan probe real-time PCR-based detection assay targeting the 16S rRNA gene of P. anguilliseptica was developed. This assay was specific for P. anguilliseptica and sensitive in detecting as little as 300 pg P. anguilliseptica genomic DNA and 2.4 × 10⁰ plasmid copie. Challenge experiments of healthy flounder with a virulent strain of P. anguilliseptica confirmed the specificity of the designed molecular probe for detection of P. anguilliseptica. This assay represents a significant approach for diagnosing P. anguilliseptica infection in olive flounder.