Comparison of Virulence Gene Identification, Ribosomal Spacer PCR, and Pulsed-Field Gel Electrophoresis for Typing of Staphylococcus aureus Strains Isolated from Cases of Subclinical Bovine Mastitis in the United States

J Clin Microbiol. 2016 Jul;54(7):1871-1876. doi: 10.1128/JCM.03282-15. Epub 2016 May 18.

Abstract

Staphylococcus aureus is one of the most important pathogens causing contagious mastitis in dairy cattle worldwide. The objectives of this study were to determine if recently described S. aureus genotype B was present among previously characterized isolates from cases of bovine intramammary infection in the United States and to compare pulsed-field gel electrophoresis (PFGE) to the combination of ribosomal spacer PCR (RS-PCR) and virulence gene identification for typing of S. aureus strains. The hypothesis was that isolates that were previously characterized as contagious would be identified as genotype B and that the results of the two strain-typing methods would be comparable. Isolates were selected from a collection of S. aureus isolates from eight dairy farms. Mammary quarter milk somatic cell count (SCC) and N-acetyl-β-d-gluconaminidase (NAGase) activity data were known and used to evaluate strain pathogenicity. RS-PCR was performed with conventional gel electrophoresis, and PCR was used for toxin gene identification. RS-PCR patterns were associated with a specific virulence gene pattern, as previously reported. Five RS-PCR banding patterns were identified. None of the isolates were characterized as genotype B. No association between RS-PCR types and milk SCC was found; however, NAGase activity was significantly higher in milk from mammary glands infected with RS-PCR banding type 1 (RSP type 1) than in milk from those infected with RSP type 2. The discriminatory power values were 1.0 and 0.46 for PFGE and RS-PCR, respectively. These data suggest that genotype B may have a limited geographic distribution and that PFGE is more discriminatory than RS-PCR performed with conventional gel electrophoresis for typing of S. aureus isolates of bovine origin.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Animals
  • Asymptomatic Infections*
  • Cattle
  • Electrophoresis, Gel, Pulsed-Field / methods
  • Female
  • Genotype*
  • Mastitis, Bovine / microbiology*
  • Mastitis, Bovine / pathology
  • Molecular Typing / methods*
  • Polymerase Chain Reaction / methods
  • Staphylococcal Infections / microbiology
  • Staphylococcal Infections / pathology
  • Staphylococcal Infections / veterinary*
  • Staphylococcus aureus / classification*
  • Staphylococcus aureus / genetics*
  • Staphylococcus aureus / isolation & purification
  • United States
  • Virulence
  • Virulence Factors / analysis
  • Virulence Factors / genetics

Substances

  • Virulence Factors

Grants and funding

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.