title: A comparison of four immunometric assays for myeloperoxidase using luminescent and colorimetric signal detection.
abstract: This communication presents four different assay systems for the determination of myeloperoxidase in body fluids. One is based on conventional chemiluminescence, two on luminescence-amplified enzyme measurement using either spiroadamantane-1,2-dioxetanes with alkaline phosphatase or luminol/peroxidase/4-iodophenol coupled with a peroxidase label. The assays covered the range 0-600 micrograms/l peroxidase. Established reference range for EDTA plasma from healthy volunteers gave an upper limit of 250 micrograms/l (95% confidence limits). Intra-assay coefficients of variation were less than 5% for all assays, as seen in compound precision profiles. Inter-assay variation was less than 10% throughout the whole concentration range. Kinetic curves for the light production were performed over a 2 hour period for the enhanced luminescence assays. Dynamic ranges of these assays were compared with the conventional colorimetric assay using 4-nitrophenyl phosphate--alkaline phosphatase. The assay was standardized using a commercially available myeloperoxidase preparation with defined enzymatic activity. The protein content was estimated and the laboratory standard compared and calibrated in mass units.
authors.affiliation: Institute of Microbiology, Czechoslovak Academy of Science, Prague, CSFR.
authors.forename: I
authors.initials: I
authors.lastname: Janda
authors.forename: H
authors.initials: H
authors.lastname: Jaensch
authors.forename: J
authors.initials: J
authors.lastname: Braun
authors.forename: W G
authors.initials: WG
authors.lastname: Wood
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives; D000469:Alkaline Phosphatase; D000818:Animals; D002417:Cattle; D003124:Colorimetry / Q000379:methods; D007118:Immunoassay / Q000379:methods; D007124:Immunoenzyme Techniques; D007462:Iodobenzenes; D008163:Luminescent Measurements; D008165:Luminol; D008832:Microchemistry / Q000379:methods; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D009195:Peroxidase / Q000097:blood*; D010545:Peroxides; D010793:Phthalazines; D011137:Polystyrenes; D012016:Reference Values
chemical_list: D007462:Iodobenzenes; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D010545:Peroxides; D010793:Phthalazines; D011137:Polystyrenes; C061523:3-(2'-spiroadamantane)-4-methoxy-4-(3''-phosphoryloxy)phenyl-1,2-dioxetane; C008644:nitrophenylphosphate; C050181:4-iodophenol; D008165:Luminol; C066306:9-(N-(4-aminobutyl)-N-ethyl)aminobenzo(f)phthalazine-1,4-(2H,3H)dione; D009195:Peroxidase; D000469:Alkaline Phosphatase; D000218:Adamantane