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

Carla Renata Arciola, Simonetta Gamberini, Davide Campoccia, Livia Visai, Pietro Speziale, Lucilla Baldassarri, Lucio Montanaro

Research output: Contribution to journalArticle

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-bio film-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.

Original languageEnglish
Pages (from-to)408-413
Number of pages6
JournalJournal of Biomedical Materials Research - Part A
Volume75
Issue number2
DOIs
Publication statusPublished - Oct 14 2005

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Staphylococcus epidermidis
Multiplex Polymerase Chain Reaction
Polymerase chain reaction
Prostheses and Implants
Genes
Biofilms
Infection
Iron-Dextran Complex
Congo Red
Gene Deletion
Orthopedics
Agar
Polysaccharides
Surveys and Questionnaires

Keywords

  • ica operon
  • Multiplex PCR
  • Prosthesis-associated infections
  • Staphylococcus epidermidis

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials

Cite this

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title = "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",
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-bio film-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.",
keywords = "ica operon, Multiplex PCR, Prosthesis-associated infections, Staphylococcus epidermidis",
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T1 - 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

AU - Arciola, Carla Renata

AU - Gamberini, Simonetta

AU - Campoccia, Davide

AU - Visai, Livia

AU - Speziale, Pietro

AU - Baldassarri, Lucilla

AU - Montanaro, Lucio

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AB - 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-bio film-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.

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