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
• Bacterial Proteins genetics   MeSH
• Bacteriological Techniques MeSH
• Biological Evolution * MeSH
• DNA, Bacterial genetics   MeSH
• Humans MeSH
• Polymerase Chain Reaction MeSH
• Polymorphism, Restriction Fragment Length MeSH
• Repetitive Sequences, Nucleic Acid genetics   MeSH
• Sequence Analysis, DNA MeSH
• Staphylococcal Infections microbiology   veterinary   MeSH
• Staphylococcus intermedius classification   genetics   isolation & purification   MeSH
• Staphylococcus classification   genetics   isolation & purification   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article 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
• Drug Resistance, Bacterial MeSH
• Bacterial Proteins analysis   genetics   MeSH
• Gerbillinae MeSH
• Cats MeSH
• Rabbits MeSH
• Micrococcal Nuclease analysis   genetics   MeSH
• Multiplex Polymerase Chain Reaction MeSH
• Animal Diseases epidemiology   microbiology   MeSH
• Polymorphism, Restriction Fragment Length MeSH
• Dogs MeSH
• RNA, Ribosomal, 16S analysis   MeSH
• Cattle MeSH
• Staphylococcal Infections epidemiology   microbiology   veterinary   MeSH
• Staphylococcal Protein A analysis   genetics   MeSH
• Staphylococcus intermedius classification   genetics   isolation & purification   pathogenicity   MeSH
• Bacterial Typing Techniques MeSH
• Body Fluids microbiology   MeSH
• Animals MeSH
• Check Tag
• Cats MeSH
• Rabbits MeSH
• Dogs MeSH
• Cattle MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Poland MeSH