Abstract

A set of degenerate PCR primers was designed and used to amplify and sequence about 75% of the catalase(kat) gene from each of 49 staphylococcal strains. In some strains of Staphylococcus xylosus, S. saprophyticus, andS. equorum, two catalase genes, katA and katB, were found. A phylogenetic tree was generated and showeddiversities among 66 partial (about 900-bp) staphylococcal kat nucleotide sequences (including 17 sequencesfound in GenBank) representing 26 different species. The topology of this tree showed a distribution ofstaphylococcal species similar, but not identical, to those reported previously based on 16S rRNA, hsp60, sodA,rpoB, tuf, and gap genes. The kat gene sequences were less conserved than those of 16S rRNA, rpoB, hsp60, andtuf genes and slightly more conserved than those of the gap gene. Therefore, kat gene sequence analysis mayprovide an additional marker for inferring phylogenetic relationships of staphylococci. Moreover, the discretenucleotide polymorphism revealed in this gene could be exploited for rapid, low-cost identification of staphylococcalspecies through PCR-restriction fragment length polymorphism (RFLP) analysis. In this study, aPCR-RFLP assay performed by using only the TaqI restriction enzyme was successfully developed for rapidunequivocal identification/differentiation, at species and subspecies levels, of coagulase-positive staphylococci(CPS). The assay was validated by testing the DNA from 100 staphylococcal strains, including reference andwild CPS strains isolated from different environments. This reliable, rapid, and low-cost approach (requiringabout 6 h from DNA isolation to the achievement of results and <5 Euros for each strain tested) allowedunambiguous identification of all the strains assayed, including the newly described S. delphini and S. pseudintermediusCPS species.

Autore Pugliese

• Fusco Vincenzina

Tutti gli autori

• Blaiotta G.; Fusco V.; Ercolini D.; Pepe O.; Coppola S.

Titolo volume/Rivista

Journal of clinical microbiology

Anno di pubblicazione

2010

ISSN

0095-1137

ISBN

Non Disponibile

Numero di citazioni Wos

Nessuna citazione

Ultimo Aggiornamento Citazioni

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Numero di citazioni Scopus

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Ultimo Aggiornamento Citazioni

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Settori ERC

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Codici ASJC

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