Identification and differentiation of Staphylococcus carnosus and Staphylococcus simulans by species-specific PCR assays of sodA genes.
The aim of this study was to design species-specific PCR assays for rapid and reliable identification and differentiation of Staphylococcus (S.) carnosus and S. simulans strains. Two different sets of primers, targeting the manganese-dependent superoxide dismutase (sodA) gene of S. carnosus and S. simulans, respectively, were designed. Species-specificity of both sets of primers was evaluated by using 93 strains, representing 26 different species of the genus Staphylococcus, 3 species of the genus Kocuria (K.), 1 species of the genus Micrococcus (Mic.) and 1 species of the genus Macrococcus (Mac.) as reference. By using primers simF and simR the expected PCR fragment was obtained only when purified DNA from S. simulans strains was used. Amplification performed by using primers carF and carR produced a PCR fragment of the expected length, when DNA from strains of S. carnosus and S. condimenti were used as template. Nevertheless, DraI digestion of the carF/carR PCR fragment allowed a clear differentiation of strains of these two species. Species-specific PCR assays designed during this study, overcoming many of the limitations of the traditional identification procedures, can be considered a valid strategy for detection and identification of S. carnosus and S. simulans strains. The rapidity (about 4h from DNA isolation to results), the reliability and low cost of the PCR procedures established suggests that the methods may be profitably applied for specific detection and identification of S. carnosus, S. condimenti and S. simulans strains in starter cultures and meat products.
Blaiotta
Giuseppe
G
Dipartimento di Scienza degli Alimenti, Sezione di Microbiologia Agraria, Alimentare, Ambientale e di Igiene, Stazione di Microbiologia Industriale, Università degli Studi di Napoli "Federico II", 80055 Portici, Italy. blaiotta@unina.it
Casaburi
Annalisa
A
Villani
Francesco
F
D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques; D017931:DNA Primers; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D013045:Species Specificity; D013210:Staphylococcus / Q000145:classification* / Q000201:enzymology / Q000235:genetics; D013482:Superoxide Dismutase / Q000235:genetics*
D001426:Bacterial Proteins; D017931:DNA Primers; C095500:SodA protein, Bacteria; D013482:Superoxide Dismutase