A group of 59 putative strains of Staphylococcus intermedius/Staphylococcus pseudintermedius deposited in the Czech National Collection of Type Cultures (CNCTC, National Institute for Public Health, Prague, Czech Republic) and the National Reference Laboratory for Staphylococci (NRL for Staphylococci, National Institute for Public Health, Prague, Czech Republic) was reclassified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). There the biggest human collection of S. pseudintermedius in Europe was analysed; 44 samples (75%) were of human origin. Twenty-two percent (n = 13) of the strains were isolated from animals, and two staphylococci were of unknown origin. This study revealed the prevalence of Staphylococcus pseudintermedius (94%, n = 53) vs. Staphylococcus intermedius (6%, n = 6) in the collection of human and veterinary staphylococci after reclassification. Results of PCR-RFLP analysis were verified by comparison with a repetitive element sequence-based polymerase chain reaction (Rep-PCR) analysis on 26 (44%) randomly selected strains. Due to a low-resolution ability of PCR-RFLP to separate Staphylococcus intermedius from Staphylococcus delphini, four isolates of Staphylococcus intermedius were biochemically verified further to exclude the presence of Staphylococcus delphini in the collection. Our results indicate that S. intermedius and S. pseudintermedius have occurred independently over an age-long period of their co-evolution.
- MeSH
- bakteriální proteiny genetika MeSH
- bakteriologické techniky MeSH
- biologická evoluce * MeSH
- DNA bakterií genetika MeSH
- lidé MeSH
- polymerázová řetězová reakce MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- repetitivní sekvence nukleových kyselin genetika MeSH
- sekvenční analýza DNA MeSH
- stafylokokové infekce mikrobiologie veterinární MeSH
- Staphylococcus intermedius klasifikace genetika izolace a purifikace MeSH
- Staphylococcus klasifikace genetika izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The aim of this study was to determine the species distribution among 44 randomly selected clinical isolates (30 mecA-positive and 14 mecA-negative) of animal origin previously identified as Staphylococcus intermedius by phenotypic tests and species-specific PCR amplification of the 16S rRNA gene. For this purpose, we used a multiplex PCR for the detection of the nuc gene and restriction fragment length polymorphism analysis of pta gene amplified by PCR. Both methods allow discrimination of Staphylococcus pseudintermedius from the other closely related members of the S. intermedius group and other coagulase-positive staphylococci isolated from animals. Genetic diversity of S. pseudintermedius strains was analyzed by staphylococcal protein A-encoding gene (spa) typing. Multiplex PCR method was used to identify staphylococcal cassette chromosome mec (SCCmec) type in mecA-positive strains. All isolates previously identified as S. intermedius were shown to belong to S. pseudintermedius. According to PCR-based SCCmec typing, SCCmecIII was the most prevalent type (n = 23), and solely seven isolates were designated as non-typeable. Furthermore, the assessment of spa-typing results revealed that the majority of all strains (n = 27) harbored spa type t02, and 17 strains were classified as non-typeable.
- MeSH
- bakteriální léková rezistence MeSH
- bakteriální proteiny analýza genetika MeSH
- Gerbillinae MeSH
- kočky MeSH
- králíci MeSH
- mikrokoková nukleasa analýza genetika MeSH
- multiplexová polymerázová řetězová reakce MeSH
- nemoci zvířat epidemiologie mikrobiologie MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- psi MeSH
- RNA ribozomální 16S analýza MeSH
- skot MeSH
- stafylokokové infekce epidemiologie mikrobiologie veterinární MeSH
- stafylokokový protein A analýza genetika MeSH
- Staphylococcus intermedius klasifikace genetika izolace a purifikace patogenita MeSH
- techniky typizace bakterií MeSH
- tělesné tekutiny mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- kočky MeSH
- králíci MeSH
- psi MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Polsko MeSH
