LOOP-MEDIATED ISOTHERMAL AMPLIFICATION - AN ASSAY FOR THE DETECTION OF ATYPICAL FURUNCULOSIS CAUSED BY AEROMONAS SALMONICIDA IN ATLANTIC COD, GADUS MORHUA

摘要

A loop-mediated isothermal amplification (LAMP)-based assay was developed for the detection of atypical furunculosis, caused by Aeromonas salmonicida in Atlantic cod, Gadus morhua. Gene gyvB encoding the B subunit of DNA gyrase present in the pathogenwas selected for designing five sets of primers targeting the flanking regions of the gene. The primers were specific for the detection of A. salmonicida with no cross reactions to other bacterial pathogens commonly infecting Atlantic cod, e.g., Vibrioanguillarum, Fran-cisella piscicida, Yersinia ruckeri and some endogenous bacteria found in the gut of Atlantic cod. The detection limit of the assay was 1 picogram of bacterial DNA mLr; whereas there was a decrease in detection limit by 1 log dilution in the presence of mucus as inhibitor Because of its specificity and sensitivity, LAMP can be considered a useful tool in routine surveillance programs in aquaculture systems for monitoring atypical furunculosis in Atlantic cod and other marine species.