<h5>Introduction</h5> As insulin resistant horses risk laminitis, 1 accurate measurement of plasma insulin concentrations is important. The Siemens Coat-A-Count Insulin Radioimmunoassay (RIA) (Siemens Medical Solutions Diagnostics, Los Angeles, CA) is widely used to quantify insulin in equine plasma. This assay is optimised to detect human insulin, with an anti-human insulin antibody, but there is good reason to anticipate cross-reactivity based on the highly conserved nature of insulin among mammalian species. 2 We evaluated this assay due to concerns that the RIA did not measure equine insulin accurately when high concentrations were determined through dilution.</P><h5>Materials & Methods</h5> The Siemens Coat-A-Count Insulin assay is a solid-phase RIA, wherein 125 I-labelled insulin competes with endogenous insulin for binding sites on an insulin-specific antibody.</P>Samples were equine plasma collected from previous studies in which high and low insulin concentrations had been observed. All samples were collected into lithium heparin tubes, kept on ice, centrifuged at 4 °C within 2 h and stored at -20 °C, until being assayed in duplicate.</P>An internal laboratory quality control (QC) sample of pooled equine plasma was assayed in duplicate at the start, middle and end of each assay, to determine intra-assay variation. Studies of dilutional parallelism and recovery
Journal of Equine Veterinary Science – Elsevier
Published: May 1, 2009
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