An improved multiple-locus variable-number of tandem repeat analysis (MLVA) for the fish pathogen Francisella noatunensis using capillary electrophoresis

BMC Veterinary Research
Samuel DuoduDuncan J Colquhoun

Abstract

Francisellosis, caused by the bacterium Francisella noatunensis subsp. noatunensis, remains a serious threat to Atlantic cod (Gadhus morhua) farming in Norway and potentially in other countries. As outbreak strains appear clonal in population structure, access to highly discriminatory typing tools is critical for understanding the epidemiology of francisellosis infections in aquaculture. In this study, a simplified multiple-locus variable-number of tandem repeat analysis (MLVA) targeting five highly polymorphic variable number of tandem repeat (VNTR) loci in a single multiplex PCR was developed to rapidly discriminate between outbreak strains. The assay resulted in identification of at least 13 different allelic profiles or subpopulations among 91 F. noatunensis isolates from farmed cod in Norway. The VNTR loci appear relatively stable, with isolates originating from individual outbreaks showing identical MLVA profiles following repeated passage. MLVA displayed greater discriminatory power than pulse-field gel electrophoresis (PFGE). Both MLVA and PFGE show good epidemiological concordance by their abilities to separate outbreak strains from epidemiologically unrelated isolates. The MLVA method presented here is robust, easy to...Continue Reading

References

Dec 24, 1998·Nucleic Acids Research·G Benson
Jun 29, 1999·Current Opinion in Microbiology·A van Belkum
Dec 15, 2006·Journal of Clinical Microbiology·Alessandro C PasqualottoMichael J Anderson
Nov 6, 2007·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·A van BelkumUNKNOWN European Society of Clinical Microbiology and Infectious Diseases (ESCMID) Study Group on Epidemiological Markers (ESGEM)
Sep 4, 2007·International Journal of Systematic and Evolutionary Microbiology·J MikalsenD J Colquhoun
Mar 10, 2011·Veterinary Research·Duncan J Colquhoun, Samuel Duodu

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Datasets Mentioned

BETA
GM2212

Methods Mentioned

BETA
PCR
electrophoresis
PCRs

Software Mentioned

LIAN
GeneMapper
BioNumerics
Primer3
MLVA
LIAN Linkage analysis

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