Multiplex Taqman Real-Time Pcr For Detecting Aeromonas Hydrophila, A. Veronii and A. Schubertii

Abstract

In this study, pathogenic Aeromonas hydrophila, A. veronii, and A. schubertii from fish were detected by multiplex TaqMan real-time PCR assay. The assay utilized three pairs of specific primers and three corresponding TaqMan probes designed to detect the aerolysin gene in A. hydrophila, the aerolysin gene in A. veronii, and the gyrB gene in A. schubertii. The specificity of the probe and primers was evaluated. The detectable concentration for the multiplex real-time PCR was 3.33×10^1 copies/μL per reaction, respectively. In addition, the coefficient of variation was less than 1.5% for both intra-and inter-assay. The assay, when screened for 120 cultured fish samples, showed 76.7% positive for Aeromonas spp. and could accurately identify these bacterial strains. These results indicated that this assay could be used as an effective tool for rapid detection and epidemiology investigation of A. hydrophila, A. veronii, and A. schubertii.