title: Design and evaluation of specific PCR primers for rapid and reliable identification of Staphylococcus xylosus strains isolated from dry fermented sausages.
abstract: Rapid and reliable identification of Staphylococcus xylosus was achieved by species-specific PCR assays. Two sets of primers, targeting on xylulokinase (xylB) and 60 kDa heat-shock protein (hsp60) genes of S. xylosus, respectively, were designed. Species-specificity of both sets of primers was evaluated by using 27 reference strains of the DSM collection, representing 23 different species of the Staphylococcus genus and 3 species of the Kocuria genus. Moreover, 90 wild strains isolated from different fermented dry sausages were included in the analysis. By using primers xylB-F and xylB-R the expected PCR fragment was obtained only when DNA from S. xylosus was used. By contrast, amplification performed by using primers xylHs-F and xylHs-R produced a single PCR fragment, of the expected length, when DNA from S. xylosus, S. haemolyticus, S. intermedius and S. kloosii were used as template. Nevertheless, AluI digestion of the xylHs-F/xylHs-R PCR fragment allowed a clear differentiation of these 4 species. The rapidity (about 4 h from DNA isolation to results) and reliability of the PCR procedures established suggests that the method may be profitably applied for specific detection and identification of S. xylosus strains.
authors.affiliation: 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", Portici, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Blaiotta
authors.forename: Carmelina
authors.initials: C
authors.lastname: Pennacchia
authors.forename: Eugenio
authors.initials: E
authors.lastname: Parente
authors.forename: Francesco
authors.initials: F
authors.lastname: Villani
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D018834:Chaperonin 60 / Q000235:genetics; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000737:chemistry / Q000302:isolation & purification; D005516:Food Microbiology; D008461:Meat Products / Q000382:microbiology*; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013210:Staphylococcus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D017853:Phosphotransferases (Alcohol Group Acceptor); C032700:xylulokinase