{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T13:51:28Z","timestamp":1792590688558,"version":"4.5.0"},"reference-count":37,"publisher":"Microbiology Society","issue":"8","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2009,8,1]]},"abstract":"\n Biofilms provide a reservoir of potentially infectious micro-organisms that are resistant to antimicrobial agents, and their importance in the failure of medical devices and chronic inflammatory conditions is increasingly being recognized. Particular research interest exists in the association of biofilms with wound infection and non-healing, i.e. chronic wounds. In this study, fluorescent\n in situ<\/jats:italic>\n hybridization (FISH) was used in combination with confocal laser scanning microscopy (CLSM) to detect and characterize the spatial distribution of biofilm-forming bacteria which predominate within human chronic skin wounds (\n Pseudomonas aeruginosa<\/jats:italic>\n ,\n Staphylococcus aureus<\/jats:italic>\n ,\n Streptococcus<\/jats:italic>\n sp. and\n Micrococcus<\/jats:italic>\n sp.).\n In vitro<\/jats:italic>\n biofilms were prepared using a constant-depth film fermenter and a reconstituted human epidermis model.\n In vivo<\/jats:italic>\n biofilms were also studied using biopsy samples from non-infected chronic venous leg ulcers. The specificity of peptide nucleic acid (PNA) probes for the target organisms was confirmed using mixed preparations of planktonic bacteria and multiplex PNA probing. Identification and location of individual bacterial species within multi-species biofilms demonstrated that\n P. aeruginosa<\/jats:italic>\n was predominant. CLSM revealed clustering of individual species within mixed-species biofilms. FISH analysis of archive chronic wound biopsy sections showed bacterial presence and allowed bacterial load to be determined. The application of this standardized procedure makes available an assay for identification of single- or multi-species bacterial populations in tissue biopsies. The technique provides a reliable tool to study bacterial biofilm formation and offers an approach to assess targeted biofilm disruption strategies\n in vivo<\/jats:italic>\n .\n <\/jats:p>","DOI":"10.1099\/mic.0.028712-0","type":"journal-article","created":{"date-parts":[[2009,7,28]],"date-time":"2009-07-28T21:51:17Z","timestamp":1248817877000},"page":"2603-2611","source":"Crossref","is-referenced-by-count":180,"title":["Detection and identification of specific bacteria in wound biofilms using peptide nucleic acid fluorescent in situ hybridization (PNA FISH)"],"prefix":"10.1099","volume":"155","author":[{"given":"Sladjana","family":"Malic","sequence":"first","affiliation":[{"name":"Tissue Engineering and Reparative Dentistry, School of Dentistry, Cardiff University, Heath Park, Cardiff CF14 4XY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katja E.","family":"Hill","sequence":"additional","affiliation":[{"name":"Tissue Engineering and Reparative Dentistry, School of Dentistry, Cardiff University, Heath Park, Cardiff CF14 4XY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anthony","family":"Hayes","sequence":"additional","affiliation":[{"name":"School of Biosciences, Cardiff University, Park Place, Cardiff CF10 3US, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven L.","family":"Percival","sequence":"additional","affiliation":[{"name":"School of Medicine, University of West Virginia, Morgantown, WV 26506, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David W.","family":"Thomas","sequence":"additional","affiliation":[{"name":"Tissue Engineering and Reparative Dentistry, School of Dentistry, Cardiff University, Heath Park, Cardiff CF14 4XY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David W.","family":"Williams","sequence":"additional","affiliation":[{"name":"Tissue Engineering and Reparative Dentistry, School of Dentistry, Cardiff University, Heath Park, Cardiff CF14 4XY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/S0958-1669(00)00204-4","article-title":"The identification of microorganisms by fluorescence in situ hybridisation","volume":"12","author":"Amann","year":"2001","journal-title":"Curr Opin Biotechnol"},{"key":"R2","doi-asserted-by":"crossref","first-page":"171","DOI":"10.12968\/jowc.2007.16.4.27025","article-title":"Bacterial profiling using skin grafting, standard culture and molecular bacteriological methods","volume":"16","author":"Andersen","year":"2007","journal-title":"J Wound Care"},{"key":"R3","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1111\/j.1524-475X.2007.00283.x","article-title":"Why chronic wounds will not heal: a novel hypothesis","volume":"16","author":"Bjarnsholt","year":"2008","journal-title":"Wound Repair Regen"},{"key":"R4","article-title":"US Patent 6664045. 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