A real-time PCR assay to detect the Panton Valentine Leukocidin toxin in staphylococci: screening Staphylococcus schleiferi subspecies coagulans strains from companion animals

Vet Microbiol. 2005 Apr 25;107(1-2):139-44. doi: 10.1016/j.vetmic.2005.01.002.

Abstract

Recent reports suggest that methicillin-resistant strains of Staphylococcus schleiferi subspecies coagulans are now commonly isolated from dogs. Given the association of a potentially mobile SCCmec type IV element with lysogenic phage-encoded Panton Valentine Leukocidin (PVL) toxin genes in community-acquired methicillin-resistant Staphylococcus aureus strains we hypothesized that methicillin-resistant S. schleiferi ssp. coagulans strains may also encode PVL toxin genes. Forty S. schleiferi ssp. coagulans strains isolated from companion animals were studied. Susceptibility to oxacillin was determined by broth microdilution and all isolates were screened by PCR for the presence of the mecA gene. SCCmec typing was performed on 14 isolates. A real-time PCR assay was developed for the detection of the PVL genes using a SmartCycler. Pulsed-field gel electrophoresis (PFGE) was performed to determine whether S. schleiferi ssp. coagulans strains were homogeneous. Twenty-eight of the 40 isolates (70%) were resistant to oxacillin and 26/28 possessed the mecA gene by PCR. SCCmec IV was identified in seven strains; the other seven isolates were not typable by this technique. All 40 strains were negative for the PVL toxin gene. PFGE showed a heterogeneous population and 13 different profiles were determined. In conclusion, this study showed that PVL toxin genes were not detected in a heterogeneous population of methicillin-resistant S. schleiferi ssp. coagulans strains isolated from companion animals.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Animals, Domestic
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Toxins
  • Coagulase / metabolism
  • Dog Diseases / drug therapy
  • Dog Diseases / microbiology*
  • Dogs
  • Drug Resistance, Bacterial
  • Electrophoresis, Gel, Pulsed-Field / veterinary
  • Exotoxins
  • Leukocidins / genetics
  • Leukocidins / isolation & purification*
  • Methicillin Resistance* / genetics
  • Microbial Sensitivity Tests / veterinary
  • Otitis Media / drug therapy
  • Otitis Media / microbiology
  • Otitis Media / veterinary
  • Oxacillin / pharmacology
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / veterinary*
  • Staphylococcal Skin Infections / drug therapy
  • Staphylococcal Skin Infections / microbiology
  • Staphylococcal Skin Infections / veterinary*
  • Staphylococcus aureus
  • Staphylococcus* / drug effects
  • Staphylococcus* / genetics
  • Staphylococcus* / isolation & purification
  • Staphylococcus* / metabolism

Substances

  • Anti-Bacterial Agents
  • Bacterial Toxins
  • Coagulase
  • Exotoxins
  • Leukocidins
  • Panton-Valentine leukocidin
  • Oxacillin