Yishuai Du
Chinese Academy of Sciences, China
Title: Development and application of a real-time PCR assay for the detection of Aeromonas salmonicida
Biography
Biography: Yishuai Du
Abstract
A rapid, economical, specific and sensitive real-time polymerase chain reaction (RT-PCR) assay coupled with SYBR Green I chemistry was developed for the quantitative detection of Aeromonas salmonicida (A. salmonicida) isolated from farmed Atlantic salmon (Salmo salar) with the symptoms of furunculosis. The set of primers designed from the virulence array protein (vapA) gene was specific to the A. salmonicida and didn’t cross-react with other bacteria. Compared with the conventional PCR, RT-PCR had a lower detection limit of 5.6 copies of the positive plasmids. The standard curve, which showed the relationship between the copies of A. salmonicida and its cycle threshold (CT) value, could be described as: log (copies of A. salmonicida) = -0.3213 CT + 10.721. The quantitative detection of copies of A. salmonicida in different tissues of the moribund Atlantic salmon showed that A. salmonicida could be detected in all tissues detected; the spleen contained the largest number of A. salmonicida, and then the kidney. These results suggest that the RT-PCR assay reported here is a specific, sensitive and quantitative method for detecting A. salmonicida in different tissues of Atlantic salmon. It can be used for the routine tests of A. salmonicida in the local aquaculture enterprise and for the research of infection routes of A. salmonicida to Atlantic salmon