Affiliation:
1. Sakhalin branch of «VNIRO» («SakhNIRO»)
2. S.P. Kapitsa Research Institute of Technology («RIT UlSU»)
3. P.A. Stolypin Ulyanovsk State Agrarian University («UlSAU»)
Abstract
The aim of the work was to select and test several pairs of primers to identify the causative agent of A. salmonicida salmon furunculosis by PCR.The methods: six bacterial cultures isolated from pre-spawning chum salmon with and without external signs of furunculosis, caught in rivers in the south of Sakhalin, were used as test cultures during the testing of primers. Ready-made kits D-Cells‑250 and Intifica TaqM master mix were used to isolate bacterial DNA and prepare PCR mixtures. Amplification was carried out in a thermocycler T‑100 ThermoCycler (Bio-Rad). Detection of PCR products was carried out by electrophoresis in 1.5% agarose gel in triacetate buffer (TAE). To view the results and document them, the Bio-Rad Gel DOC XR+system was used.The results: the species specificity of three pairs of primers selected for the identification of the causative agent of furunculosis by PCR was experimentally confirmed. Testing of the selected primers was carried out on bacterial cultures isolated from pre-spawning chum salmon individuals with and without manifestations of furunculosis. According to the results of testing, all bacterial isolates were assigned to the species A. salmonicida.Novelty of the work: for the first time, a comparison of primers proposed by several authors for the identification of the causative agent of furunculosis was performed on bacterial cultures isolated from salmon of the Far Eastern region.Practical significance: the obtained results were used to prepare methodological guidelines for the identification of the causative agent of salmon furunculosis A. salmonicida by PCR, which reduces the time of diagnosis of the disease.
Publisher
Russian Federal Research Institute of Fisheries and Oceanography
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