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Molecular Characterization of Rhodococcus equi from Horse-Breeding Farms by Means of Multiplex PCR for the vap Gene Family

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Abstract

This study evaluated the molecular characteristics of Rhodococcus equi isolates obtained from horses by a multiplex PCR assay that amplifies the vap gene family (vapA, -B, -C, -D, -E, -F, -G, and -H). A total of 180 R. equi isolates were studied from four different sources, namely healthy horse feces (112), soil (12), stalls (23), and clinical isolates (33) from horse-breeding farms. The technique was performed and confirmed by sequencing of amplified vap gene family controls. Thirty-two (17.8%) of the R. equi isolates were positive for the vapA gene and carried at least three other vap genes. All 147 isolates from equine feces, stalls, and soil failed to demonstrate any genes associated with virulence-inducing proteins. About 32 (97.0%) out of the 33 clinical equine isolates tested positive for the multiplex PCR assay for the vap gene family. They demonstrated six molecular profiles: 100% featured the vapA, vapD, and vapG genes, 86.6% vapF, 76.6% vapH, 43.3% vapC, 36.6% vapE, and none vapB. The most frequent molecular profile was vap A, -D, -F, G, and -H, where this profile was present in 37.5% of the strains. Moreover, there was no molecular epidemiological pattern for R. equi isolates that uniquely mapped to each horse-breeding farm studied. Our proposed technique allows the identification of eight members of the vap gene family (vapA, B, -C, -D, -E, -F, -G, and -H). It is a practical and efficient method of conducting clinical and epidemiological studies on R. equi isolates.

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Acknowledgments

This research was funded by the Conselho Nacional de Pesquisa e Desenvolvimento Tecnológico, CNPq/Brazil grant 471694/2007-0 and Fundação de Apoio a Pesquisa do Estado do Rio Grande do Sul, Fapergs/Brazil grant 07/50917.0.

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Authors and Affiliations

  1. Medicina Veteirnaria Preventiva, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil

    Fernanda Monego & Franciele Maboni

  2. Universidade Federal do Vale do São Francisco, Petrolina, PE, Brazil

    Cristina Krewer

  3. Veterinary, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil

    Agueda Vargas

  4. Veterinary, Universidade Federal do Vale do São Francisco, Petrolina, PE, Brazil

    Mateus Costa

  5. Biologia, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil

    Elgion Loreto

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  1. Fernanda Monego
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  2. Franciele Maboni
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  3. Cristina Krewer
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  4. Agueda Vargas
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  5. Mateus Costa
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  6. Elgion Loreto
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Correspondence to Fernanda Monego.

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Monego, F., Maboni, F., Krewer, C. et al. Molecular Characterization of Rhodococcus equi from Horse-Breeding Farms by Means of Multiplex PCR for the vap Gene Family. Curr Microbiol 58, 399–403 (2009). https://doi.org/10.1007/s00284-009-9370-6

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  • DOI: https://doi.org/10.1007/s00284-009-9370-6

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