title: Effects of sample handling on adrenocorticotropin concentration measured in canine plasma, using a commercially available radioimmunoassay kit.
abstract: A commercially available radioimmunoassay (RIA) kit for measurement of human adrenocorticotropin (hACTH) was validated for use in dogs. Assay sensitivity was 3 pg/ml. Intra-assay coefficient of variation (x 100; CV) for 3 canine plasma pools was 3.0 (mean +/- SD, 33 +/- 0.99 pg/ml), 4.2 (71 +/- 2.4 pg/ml) and 3.7 (145 +/- 3.7 pg/ml) %. Interassay CV for 2 plasma pools measured in 6 assays was 9.8 (37 +/- 3.6 pg/ml) and 4.4 (76 +/- 3.4 pg/ml) %, respectively. Dilutional parallelism was documented by assaying 2 pools of canine plasma at 3 dilutions and correcting the measured result for dilution. Corrected mean concentrations for the first pool were 33 (+/- 0.99), 36 (+/- 4.3), and 33 (+/- 6.8) pg/ml; corrected mean concentrations for the second pool were 145 (+/- 5.4), 141 (+/- 10.8) and 125 (+/- 3.4) pg/ml. Recovery of 1-39hACTH added to canine plasma (6.25, 12.5, 25.0, 50.0, and 100.0 pg/ml) was linear and quantitative (slope = 0.890, R2 = 0.961). To test whether anticoagulant or the protease inhibitor, aprotinin, influences ACTH concentration in canine plasma, ACTH was measured in canine blood collected in 4 tubes containing anticoagulant: heparin (H), heparin + 500 kallikrein inhibitor units (KIU) of aprotinin/ml (HA), EDTA (E), and EDTA + aprotinin (EA). Plasma ACTH concentration was the same when samples containing H and HA, or HA and E were compared, and was significantly (P less than 0.01) lower in samples containing EA.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul 55108.
authors.forename: R L
authors.initials: RL
authors.lastname: Hegstad
authors.forename: S D
authors.initials: SD
authors.lastname: Johnston
authors.forename: D M
authors.initials: DM
authors.lastname: Pasternak
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000097:blood*; D000818:Animals; D007611:Aprotinin / Q000494:pharmacology; D004285:Dogs; D004492:Edetic Acid; D006493:Heparin / Q000494:pharmacology; D011863:Radioimmunoassay / Q000662:veterinary; D011933:Reagent Kits, Diagnostic / Q000662:veterinary; D013048:Specimen Handling / Q000379:methods / Q000662:veterinary*; D013997:Time Factors
chemical_list: D011933:Reagent Kits, Diagnostic; D000324:Adrenocorticotropic Hormone; D006493:Heparin; D007611:Aprotinin; D004492:Edetic Acid