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Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR RS-PCR
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Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR RS-PCR

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Some coagulase negative Staphylococcus spp. isolated from buffalo can be misidentified as Staphylococcus aureus by

Camila C de Almeida1,2, Lucas J L Pizauro3,2, Glenn A Soltes2

  • 1Agriculture and Livestock Microbiology Graduation Program, Department of Veterinary Pathology, São Paulo State University (Unesp), School of Agricultural and Veterinarian Sciences, Jaboticabal, Brazil.

BMC Research Notes
|June 1, 2018
PubMed
Abstract

Insights

Conventional methods may overestimate Staphylococcus aureus in buffalo mastitis. Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) and PCR gene targets offer more accurate identification of S. aureus.

Area of Science:

  • Veterinary Microbiology
  • Bovine Mastitis Research
  • Diagnostic Microbiology

Background:

  • Staphylococcus aureus is a primary cause of buffalo mastitis.
  • Its prevalence might be overestimated by traditional identification methods.

Purpose of the Study:

  • To compare conventional phenotypic and genotypic assays with MALDI-TOF MS and real-time PCR for S. aureus identification.
  • To evaluate novel PCR targets (cydB and coa genes) for S. aureus detection.

Main Methods:

  • Milk and environmental samples were analyzed using conventional methods (Baird Parker agar, catalase, plasma clotting, Sa442 PCR).
  • Samples were further tested with latex agglutination, MALDI-TOF MS, 16S rRNA gene sequencing, and real-time PCR for cydB and coa genes.

Main Results:

  • Out of 32 putative S. aureus strains, only 8 were confirmed by MALDI-TOF MS and 16S rRNA sequencing.
  • MALDI-TOF MS identified other Staphylococcus species, including S. chromogenes and S. agnetis.
  • S. aureus-specific cydB PCR confirmed all 8 S. aureus strains; coa PCR detected 7/8 S. aureus and some other species.

Conclusions:

  • MALDI-TOF MS and molecular methods provide more accurate identification of S. aureus in buffalo mastitis cases.
  • Conventional methods may lead to overestimation of S. aureus prevalence.
  • Novel PCR targets show potential for specific S. aureus detection.
Keywords:
MastitisSpecies-specific PCR testsStaphylococcus aureuscydB PCR

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