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A multiplex PCR method for the detection of all five individual genes of ica locus in Staphylococcus epidermidis . A survey on 400 clinical isolates from prosthesis‐associated infections
Author(s) -
Arciola Carla Renata,
Gamberini Simonetta,
Campoccia Davide,
Visai Livia,
Speziale Pietro,
Baldassarri Lucilla,
Montanaro Lucio
Publication year - 2005
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.30445
Subject(s) - staphylococcus epidermidis , biofilm , multiplex polymerase chain reaction , biology , locus (genetics) , microbiology and biotechnology , gene , staphylococcus , polymerase chain reaction , multiplex , staphylococcus aureus , genetics , bacteria
Abstract In Staphylococcus epidermidis , ica locus encodes for the synthesis of a polysaccharide intercellular adhesin (slime or biofilm). A multiplex polymerase chain reaction (PCR) for the detection of the five individual genes of ica locus was developed, with the aim to probe the set of genes in a large collection of Staphylococcus epidermidis clinical isolates. Single representative fragments for icaR , icaA , icaD , icaB , and icaC genes were selected. Multiplex PCR was applied to two reference Staphylococcus epidermidis strains [the non–biofilm‐forming ATCC 12228 and the biofilm‐forming ATCC 35984 (RP62A)] and to 400 clinical isolates of Staphylococcus epidermidis from orthopedic prosthesis associated infections. The gene profile was compared with the phenotypic biofilm‐forming ability, evaluated by means of an optimized Congo red agar (CRA) plate test. Among the clinical isolates, 228 (57%) turned out completely ica positive and were biofilm producing. Among the 172 non–biofilm‐forming strains (43%), 164 (41%) were completely ica negative and 8 strains (2%) harbored all five ica genes. The ica locus thus proves to be a cluster of strictly linked genes, without any evidence of single gene deletion. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005