title: Rapid duplex immunoassay for wound biomarkers at the point-of-care.
abstract: In this study we describe a novel method of sampling and quantifying wound biomarkers for clinical settings. We believe the chosen format will allow rapid assessments of wound healing and provide biomarker evidence-based decision points for treatment of the wound at the time of presentation. The wound monitoring principle uses a proprietary sample collection tool (a thermally reversible hydrogel) to sample and isolate biomarkers within a wound environment without further sample extraction/preparation steps. We show how gel samples can be analysed in a lateral flow assay format utilising fluorescent microspheres with optically discrete emission characteristics and demonstrate quantitative detection of two analytes (duplexing) achieved in a single test line. As a model assay, the chronic wound biomarkers interleukin 6 (IL6) and tumour necrosis factor alpha (TNFα) are used. Limits of detection of 48.5 pg/mL and 55.5 pg/mL respectively in hydrogel samples and 7.15 pg/mL and 10.7 pg/mL respectively in plasma are reported. We believe this is the first literature example of quantitative detection of multiple analytes within a single test line using spectral separation to distinguish the analytes.
authors.affiliation: Biotechnology Group, Quality of Life Division, National Physical Laboratory, Teddington, Middlesex, UK. Graham.Worsley@NPL.co.uk
authors.forename: G J
authors.initials: GJ
authors.lastname: Worsley
authors.forename: S L
authors.initials: SL
authors.lastname: Attree
authors.forename: J E
authors.initials: JE
authors.lastname: Noble
authors.forename: A M
authors.initials: AM
authors.lastname: Horgan
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D015415:Biomarkers / Q000097:blood; D015374:Biosensing Techniques / Q000379:methods*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D007118:Immunoassay / Q000379:methods; D015850:Interleukin-6 / Q000097:blood*; D014409:Tumor Necrosis Factor-alpha / Q000097:blood*; D014947:Wounds and Injuries / Q000097:blood*
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate