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Molecular diagnosis of francisellosis, a systemic granulomatous inflammatory disease in Atlantic cod, Gadus morhua L

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Abstract

A PCR-based assay for the detection of Francisella noatunensis causing francisellosis in Atlantic cod, Gadus morhua has been developed. Seven sets of primers targeting the flanking regions of the genes (rpoA, sdhA, atpA, rpoB, pgm, groEL and 16S rRNA) of the pathogen were designed. Among the primers, groEL was found to be the most suitable gene candidate for detecting the pathogen, due to its high sensitivity at various annealing temperatures and specificity in detection. The detection limit of the assay was 100 pg of bacterial DNA per milliliter or 100 fg bacterial DNA (approximately 50 genome equivalents) per PCR reaction, however, the sensitivity of the reaction decreased by 1 log dilution in the presence of 1 mg mL−1 of serum and mucus samples as inhibitors. Nevertheless, the assay can potentially be used as a direct and non-lethal method to detect the pathogen in fish. Thus this PCR assay is a specific and sensitive molecular method to diagnose francisellosis in Atlantic cod, and will be helpful for controlling the infection through prompt detection of the disease in farms.

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Acknowledgements

This study is part of the Master’s thesis of the first author (A. Kulkarni) and was supported in part by the research project, “Preventive Health Care in Farmed Fish” (176528/V10) funded by the Research Council of Norway. The authors express their gratitude to Professor Are Nylund and Karl Fredrik Ottem of the University of Bergen and Professor Øivind Bergh of the Institute of Marine Research, Norway for providing bacterial samples.

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  1. Faculty of Biosciences and Aquaculture, Bodø University College, Bodø, 8049, Norway

    Amod Kulkarni, Christopher Marlowe A. Caipang, Kjetil Korsnes, Monica F. Brinchmann & Viswanath Kiron

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  1. Amod Kulkarni
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  2. Christopher Marlowe A. Caipang
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  3. Kjetil Korsnes
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  4. Monica F. Brinchmann
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  5. Viswanath Kiron
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Correspondence to Christopher Marlowe A. Caipang.

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Kulkarni, A., Caipang, C.M.A., Korsnes, K. et al. Molecular diagnosis of francisellosis, a systemic granulomatous inflammatory disease in Atlantic cod, Gadus morhua L. Vet Res Commun 35, 67–77 (2011). https://doi.org/10.1007/s11259-010-9451-9

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