Double Capture Amplification System for Bacterial Pathogen Detection in

Bacterial pathogens in salmonids, such as Renibacterium salmoninarum and Aeromonas salmonicida, are difficult to identify by conventional diagnosis due to their extremely slow growth and the absence of an efficient selective medium. Diseases caused by these pathogens represent a serious problem both in the fishing industry and for conservation efforts. While several PCR-based assays appear to be useful, they are generally complex laboratory procedures and not suitable for large scale screening. We have proposed to develop a novel screening method, the ¿Double Capture Amplification System¿ (DCAS), to detect R. salmoninarum in salmonids. This procedure combines several powerful molecular biology tools in an innovative manner to develop a rapid, non-lethal, highly sensitive assay kit. Our Phase 1 research has demonstrated the feasibility of the proposed assay and initiated development of the prototype diagnostic kit. The major goals for the second phase of this project are to adapt, and optimize the assay for the detection of Renibacterium salmoninarum in various tissues in fish, determine the sensitivity and specificity of the assay, and begin production and quality control testing of the survey kits. In addition, we will initiate studies for the adaptation of DCAS to other organisms such as Aeromonas salmonicida.