Development and Application of a Quantitative Real‐time Polymerase Chain Reaction Assay for the Detection of Aeromonas salmonicida

• Published in: Journal of the World Aquaculture Society
• Main Authors: Du, Yishuai, Liu, Ying, Xiao, Peng, Meng, Lingjie, Liu, Pengfei
• Other Authors: China Postdoctoral Science Foundation
• Format: Article in Journal/Newspaper
• Language: English
• Published: Wiley 2016
• Subjects: Agronomy and Crop Science; Aquatic Science; Atlantic salmon; Salmo salar
• Online Access: http://dx.doi.org/10.1111/jwas.12395; https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fjwas.12395; https://onlinelibrary.wiley.com/doi/pdf/10.1111/jwas.12395

Abstract A rapid, economical, specific, and sensitive quantitative real‐time polymerase chain reaction (qPCR) assay coupled with SYBR Green I chemistry was developed for the quantitative detection of Aeromonas salmonicida 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 A. salmonicida . Compared with the conventional PCR, qPCR 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 quantification cycle ( C q ) value, could be described as follows: log (copies of A. salmonicida ) = −0.3213 C q + 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; the spleen contained the largest number of A. salmonicida and then the kidney. These results suggest that the qPCR assay reported here is a specific, sensitive, and quantitative method for detecting A. salmonicida . It can be used for the routine tests of A. salmonicida in local aquaculture enterprise and for the research of infection routes of A. salmonicida to Atlantic salmon.