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A multiplex PCR to identify porcine mycoplasmas present in broth cultures

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Abstract
Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma flocculare can be present in the lungs of pigs at the same time. These three mycoplasma species all require similar growth conditions and can be recovered from clinical samples using the same media. We have developed a multiplex PCR as a helpful tool for rapid differentiation of these three species in the course of isolation. Based on the 16S ribosomal DNA sequences, three different forward primers and a single reverse primer were selected. Each forward primer was compared to available mycoplasma sequences, showing the primers to be specific. The three amplification products observed of 1129 bp (M. hyorhinis), 1000 bp (M. hyopneumoniae) and 754 bp (M. flocculare) were clearly distinguishable on a 1% agarose gel. In addition, no cross-reaction with Mycoplasma hyosynoviae, another porcine mycoplasma, was noted. This multiplex PCR using the proposed set of primers is the first reported assay that allows the simultaneous identification of the different Mycoplasma species isolated from the lungs of pigs.
Keywords
16S, 16S ribosome DNA, DIFFERENTIATION, HYORHINIS, FLOCCULARE, AMPLIFICATION, PIGS, HYOPNEUMONIAE, RESPIRATORY-TRACT, RIBOSOMAL-RNA GENE, POLYMERASE-CHAIN-REACTION, swine, PCR, M. hyorhinis, M. hyopneumoniae, M. flocculare, mycoplasma, multiplex, isolation

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Chicago
Stakenborg, Tim, Jo Vicca, Patrick Butaye, H Imberechts, Johan Peeters, Aart de Kruif, Freddy Haesebrouck, and Dominiek Maes. 2006. “A Multiplex PCR to Identify Porcine Mycoplasmas Present in Broth Cultures.” Veterinary Research Communications 30 (3): 239–247.
APA
Stakenborg, T., Vicca, J., Butaye, P., Imberechts, H., Peeters, J., de Kruif, A., Haesebrouck, F., et al. (2006). A multiplex PCR to identify porcine mycoplasmas present in broth cultures. VETERINARY RESEARCH COMMUNICATIONS, 30(3), 239–247.
Vancouver
1.
Stakenborg T, Vicca J, Butaye P, Imberechts H, Peeters J, de Kruif A, et al. A multiplex PCR to identify porcine mycoplasmas present in broth cultures. VETERINARY RESEARCH COMMUNICATIONS. 2006;30(3):239–47.
MLA
Stakenborg, Tim, Jo Vicca, Patrick Butaye, et al. “A Multiplex PCR to Identify Porcine Mycoplasmas Present in Broth Cultures.” VETERINARY RESEARCH COMMUNICATIONS 30.3 (2006): 239–247. Print.
@article{350910,
  abstract     = {Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma flocculare can be present in the lungs of pigs at the same time. These three mycoplasma species all require similar growth conditions and can be recovered from clinical samples using the same media. We have developed a multiplex PCR as a helpful tool for rapid differentiation of these three species in the course of isolation. Based on the 16S ribosomal DNA sequences, three different forward primers and a single reverse primer were selected. Each forward primer was compared to available mycoplasma sequences, showing the primers to be specific. The three amplification products observed of 1129 bp (M. hyorhinis), 1000 bp (M. hyopneumoniae) and 754 bp (M. flocculare) were clearly distinguishable on a 1\% agarose gel. In addition, no cross-reaction with Mycoplasma hyosynoviae, another porcine mycoplasma, was noted. This multiplex PCR using the proposed set of primers is the first reported assay that allows the simultaneous identification of the different Mycoplasma species isolated from the lungs of pigs.},
  author       = {Stakenborg, Tim and Vicca, Jo and Butaye, Patrick and Imberechts, H and Peeters, Johan and de Kruif, Aart and Haesebrouck, Freddy and Maes, Dominiek},
  issn         = {0165-7380},
  journal      = {VETERINARY RESEARCH COMMUNICATIONS},
  language     = {eng},
  number       = {3},
  pages        = {239--247},
  title        = {A multiplex PCR to identify porcine mycoplasmas present in broth cultures},
  url          = {http://dx.doi.org/10.1007/s11259-006-3226-3},
  volume       = {30},
  year         = {2006},
}

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