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Isolation and molecular characterization of Brucella isolates in cattle milk in Uganda.

Mugizi DR, Muradrasoli S, Boqvist S, Erume J, Nasinyama GW, Waiswa C, Mboowa G, Klint M, Magnusson U - Biomed Res Int (2015)

Bottom Line: Brucellosis is endemic in livestock and humans in Uganda and its transmission involves a multitude of risk factors like consumption of milk from infected cattle.Brucella abortus without a biovar designation was isolated from eleven out of 207 milk samples from cattle in Uganda.These isolates had a genomic monomorphism at 16 variable number tandem repeat (VNTR) loci and showed in turn high levels of genetic variation when compared with other African strains or other B. abortus biovars from other parts of the world.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Animal Resources and Bio-Security, Makerere University, P.O. Box 7062, Kampala, Uganda.

ABSTRACT
Brucellosis is endemic in livestock and humans in Uganda and its transmission involves a multitude of risk factors like consumption of milk from infected cattle. To shed new light on the epidemiology of brucellosis in Uganda the present study used phenotypic and molecular approaches to delineate the Brucella species, biovars, and genotypes shed in cattle milk. Brucella abortus without a biovar designation was isolated from eleven out of 207 milk samples from cattle in Uganda. These isolates had a genomic monomorphism at 16 variable number tandem repeat (VNTR) loci and showed in turn high levels of genetic variation when compared with other African strains or other B. abortus biovars from other parts of the world. This study further highlights the usefulness of MLVA as an epidemiological tool for investigation of Brucella infections.

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Related in: MedlinePlus

Triplex real-time PCR amplification pattern using the Brucella genus probe. Fluorescence ratio is plotted against the number of PCR cycles to monitor amplification in real-time mode. Isolates with weak Ct values (29.18 and above) had Brucella-like phenotypic characteristics and were included in this assay.
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Related In: Results  -  Collection


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fig1: Triplex real-time PCR amplification pattern using the Brucella genus probe. Fluorescence ratio is plotted against the number of PCR cycles to monitor amplification in real-time mode. Isolates with weak Ct values (29.18 and above) had Brucella-like phenotypic characteristics and were included in this assay.

Mentions: Genotyping was performed using the Multiple Locus Variable Number Tandem Repeat Analysis (MLVA), using the 16-primer-pair assay [17, 24–26]. The oligonucleotide primer pairs incorporated in the Brucella MLVA 16 assay target both the conserved and highly discriminatory regions of the Brucella genome. Each sample was run on three prescribed MLVA panels. Panel 1 consisted of moderately polymorphic minisatellite primer pairs targeting the highly conserved genomic regions of different Brucella species (bruce 06, bruce 08, bruce 11, bruce 12, bruce 42, bruce 43, bruce 45, and bruce 55). Panel 2A (bruce 18, bruce 19, and bruce 21) and panel 2B (bruce 04, bruce 07, bruce 09, bruce 16, and bruce 30) consisted of microsatellite primer pairs targeting the discriminatory genomic regions. The PCR amplification and genotyping were done according to le Flèche et al. [17] with only a modification in the total reaction volume to 30 μL. At each run, B. suis reference strain REF. 1330 (from Bruce-ladder kit) was included as shown in Figure 1.


Isolation and molecular characterization of Brucella isolates in cattle milk in Uganda.

Mugizi DR, Muradrasoli S, Boqvist S, Erume J, Nasinyama GW, Waiswa C, Mboowa G, Klint M, Magnusson U - Biomed Res Int (2015)

Triplex real-time PCR amplification pattern using the Brucella genus probe. Fluorescence ratio is plotted against the number of PCR cycles to monitor amplification in real-time mode. Isolates with weak Ct values (29.18 and above) had Brucella-like phenotypic characteristics and were included in this assay.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4352423&req=5

fig1: Triplex real-time PCR amplification pattern using the Brucella genus probe. Fluorescence ratio is plotted against the number of PCR cycles to monitor amplification in real-time mode. Isolates with weak Ct values (29.18 and above) had Brucella-like phenotypic characteristics and were included in this assay.
Mentions: Genotyping was performed using the Multiple Locus Variable Number Tandem Repeat Analysis (MLVA), using the 16-primer-pair assay [17, 24–26]. The oligonucleotide primer pairs incorporated in the Brucella MLVA 16 assay target both the conserved and highly discriminatory regions of the Brucella genome. Each sample was run on three prescribed MLVA panels. Panel 1 consisted of moderately polymorphic minisatellite primer pairs targeting the highly conserved genomic regions of different Brucella species (bruce 06, bruce 08, bruce 11, bruce 12, bruce 42, bruce 43, bruce 45, and bruce 55). Panel 2A (bruce 18, bruce 19, and bruce 21) and panel 2B (bruce 04, bruce 07, bruce 09, bruce 16, and bruce 30) consisted of microsatellite primer pairs targeting the discriminatory genomic regions. The PCR amplification and genotyping were done according to le Flèche et al. [17] with only a modification in the total reaction volume to 30 μL. At each run, B. suis reference strain REF. 1330 (from Bruce-ladder kit) was included as shown in Figure 1.

Bottom Line: Brucellosis is endemic in livestock and humans in Uganda and its transmission involves a multitude of risk factors like consumption of milk from infected cattle.Brucella abortus without a biovar designation was isolated from eleven out of 207 milk samples from cattle in Uganda.These isolates had a genomic monomorphism at 16 variable number tandem repeat (VNTR) loci and showed in turn high levels of genetic variation when compared with other African strains or other B. abortus biovars from other parts of the world.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Animal Resources and Bio-Security, Makerere University, P.O. Box 7062, Kampala, Uganda.

ABSTRACT
Brucellosis is endemic in livestock and humans in Uganda and its transmission involves a multitude of risk factors like consumption of milk from infected cattle. To shed new light on the epidemiology of brucellosis in Uganda the present study used phenotypic and molecular approaches to delineate the Brucella species, biovars, and genotypes shed in cattle milk. Brucella abortus without a biovar designation was isolated from eleven out of 207 milk samples from cattle in Uganda. These isolates had a genomic monomorphism at 16 variable number tandem repeat (VNTR) loci and showed in turn high levels of genetic variation when compared with other African strains or other B. abortus biovars from other parts of the world. This study further highlights the usefulness of MLVA as an epidemiological tool for investigation of Brucella infections.

Show MeSH
Related in: MedlinePlus