title: Brucellosis outbreak in a Swedish kennel in 2013: determination of genetic markers for source tracing.
abstract: Brucellosis is a highly infectious zoonotic disease but rare in Sweden. Nonetheless, an outbreak of canine brucellosis caused by an infected dog imported to Sweden was verified in 2013. In total 25 dogs were tested at least duplicated by the following approaches: real-time PCR for the detection of Brucella canis, a Brucella genus-specific real-time PCR, selective cultivation, and microscopic examination. The whole genome of B. canis strain SVA13 was analysed regarding genetic markers for epidemiological examination. The genome of an intact prophage of Roseobacter was detected in B. canis strain SVA13 with whole genome sequence prophage analysis (WGS-PA). It was shown that the prophage gene content in the American, African and European isolates differs remarkably from the Asian strains. The prophage sequences in Brucella may therefore serve of use as genetic markers in epidemiological investigations. Phage DNA fragments were also detected in clustered, regularly interspaced short palindromic repeats (CRISPR) in the genome of strain SVA13. In addition to the recommendations for genetic markers in Brucella outbreak tracing, our paper reports a validated two-step stand-alone real-time PCR for the detection of B. canis and its first successful use in an outbreak investigation.
authors.affiliation: National Veterinary Institute, Ulls väg 2B, 75189 Uppsala, Sweden; Swedish Forum for Biopreparedness Diagnostics (FBD), Umeå, Uppsala and Solna, Sweden; Swedish Joint Laboratory for Food Safety and Biopreparedness, Uppsala, Sweden; Uppsala University, Department of Medical Sciences, Dag Hammarskjöldsväg 17, 75185 Uppsala, Sweden. Electronic address: rene.kaden@akademiska.se.
authors.forename: Rene
authors.initials: R
authors.lastname: Kaden
authors.affiliation: National Veterinary Institute, Ulls väg 2B, 75189 Uppsala, Sweden.
authors.forename: Joakim
authors.initials: J
authors.lastname: Ågren
authors.affiliation: National Veterinary Institute, Ulls väg 2B, 75189 Uppsala, Sweden; Swedish Forum for Biopreparedness Diagnostics (FBD), Umeå, Uppsala and Solna, Sweden.
authors.forename: Viveca
authors.initials: V
authors.lastname: Båverud
authors.affiliation: National Veterinary Institute, Ulls väg 2B, 75189 Uppsala, Sweden.
authors.forename: Gunilla
authors.initials: G
authors.lastname: Hallgren
authors.affiliation: National Veterinary Institute, Ulls väg 2B, 75189 Uppsala, Sweden; Swedish Forum for Biopreparedness Diagnostics (FBD), Umeå, Uppsala and Solna, Sweden; Swedish Joint Laboratory for Food Safety and Biopreparedness, Uppsala, Sweden.
authors.forename: Sevinc
authors.initials: S
authors.lastname: Ferrari
authors.affiliation: National Veterinary Institute, Ulls väg 2B, 75189 Uppsala, Sweden.
authors.forename: Joann
authors.initials: J
authors.lastname: Börjesson
authors.affiliation: National Veterinary Institute, Ulls väg 2B, 75189 Uppsala, Sweden; Swedish Forum for Biopreparedness Diagnostics (FBD), Umeå, Uppsala and Solna, Sweden; Swedish Joint Laboratory for Food Safety and Biopreparedness, Uppsala, Sweden; National Food Agency, Hamnesplanaden 5, 75319 Uppsala, Sweden.
authors.forename: Martina
authors.initials: M
authors.lastname: Lindberg
authors.affiliation: Swedish Forum for Biopreparedness Diagnostics (FBD), Umeå, Uppsala and Solna, Sweden; Swedish Defence Research Agency, Cementvägen 20, 16490 Umeå, Sweden.
authors.forename: Stina
authors.initials: S
authors.lastname: Bäckman
authors.affiliation: Swedish Forum for Biopreparedness Diagnostics (FBD), Umeå, Uppsala and Solna, Sweden; Public Health Agency of Sweden, Tomtebodavägen 12B, 17182 Solna, Sweden.
authors.forename: Tara
authors.initials: T
authors.lastname: Wahab
mesh_terms: D000818:Animals; D001483:Base Sequence; D041002:Brucella canis / Q000235:genetics* / Q000302:isolation & purification; D002006:Brucellosis / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D004196:Disease Outbreaks / Q000662:veterinary*; D004283:Dog Diseases / Q000453:epidemiology* / Q000382:microbiology; D004285:Dogs; D005260:Female; D005819:Genetic Markers / Q000235:genetics; D016680:Genome, Bacterial / Q000235:genetics*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D017422:Sequence Analysis, DNA / Q000662:veterinary; D013045:Species Specificity; D013548:Sweden / Q000453:epidemiology; D015047:Zoonoses
chemical_list: D005819:Genetic Markers
keywords: Brucella canis; Brucellosis; Clustered-regularly-interspaced short palindromic repeats (CRISPR); Source tracing; Whole genome sequencing phage analysis (WGS-PA); Zoonosis