Evaluation of a commercially available radioimmunoassay and species-specific ELISAs for measurement of high concentrations of insulin in equine serum

Borer-Weir, Kate; Bailey, S. R.; Menzies‐Gow, N. J.; Harris, Patricia A.; Elliott, Jonathan

doi:10.2460/ajvr.73.10.1596

Cited by 34 publications

(45 citation statements)

References 18 publications

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“…for the immunoassay validation that are consistent with previously reported values using the ELISA [13]. Clinicians should therefore be aware of the limitations of the available insulin assays when interpreting results.…”

Section: Accepted Articlesupporting

confidence: 68%

“…Dilutional parallelism was assessed by diluting each of 4 high endogenous samples (101-178 µIU/ml) to 50%, 25% and 12.5% with charcoal stripped equine serum (CSS) [13]. Spike recovery was assessed by spiking 6 samples of CSS with the kit manufacturer's human insulin standard (expected values 21-175 µIU/ml).…”

Section: Accepted Articlementioning

confidence: 99%

“…All study samples were analysed in duplicate and the CV of duplicate measurements (2.6%) was good. A previously validated radioimmunoassay [13,25] has been discontinued. Alternative assays include an equine specific ELISA [13,25] and a chemiluminescent immunoassay [26].…”

Section: Accepted Articlementioning

confidence: 99%

“…A previously validated radioimmunoassay [13,25] has been discontinued. Alternative assays include an equine specific ELISA [13,25] and a chemiluminescent immunoassay [26]. Inter-assay CVs of 4-14% are reported for the chemiluminescent assay [26]; for the ELISA intra assay CVs of 8% and 11% and inter assay CVs of 7%…”

Section: Accepted Articlementioning

confidence: 99%

“…Basic validation of the radioimmunoassay e was performed as the previously validated radioimmunoassay [13] was no longer available. Briefly, intra and inter-assay CVs [14] were estimated using 7 (10-150 µIU/ml) and 15 (12-249 µIU/ml) samples respectively, each analysed two…”

Section: Sample Processing and Analysismentioning

confidence: 99%

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Use of the oral sugar test in ponies when performed with or without prior fasting

Knowles

Harris²,

Elliott

et al. 2016

Equine Veterinary Journal

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Summary Background It is recommended that the oral sugar test (OST) for insulin dysregulation (ID) be performed after an overnight fast, but fasting is impractical in ponies kept solely at pasture. There are few data on OST repeatability and reliability in ponies. Objectives To report 1) whether OST results obtained in the morning after an overnight fast or without fasting in the afternoon (FASTING/FED) can be used interchangeably, 2) time of highest insulin concentration Tmax[insulin], repeatability and reliability of insulin response to the OST when FASTING or FED and 3) dichotomous agreement (ID/normal) within a small sample when FASTING or FED. Study design Method comparison study. Methods Oral sugar tests were performed on four occasions in 10 adult native British ponies, twice FASTING and twice FED. Insulin concentrations were measured by radioimmunoassay at 0–120 min (T0,30,60,75,90,120). Differences between FASTING and FED results were assessed using mixed effects models. Indices of repeatability and reliability were calculated; dichotomous agreement was reported using kappa statistics. Results Serum [insulin] was significantly (P≤0.05) higher at T60–T90 with prior fasting (estimated differences [95% confidence intervals]): T60: 23.5 μiu/ml (8.7–38.4 μiu/ml), T75: 27.1 μiu/ml (12.3–41.8 μiu/ml), T90: 15.1 (0.36–29.9 μiu/ml). Most frequently, Tmax[ins] occurred at T30. At any single time point, within‐subject coefficients of variation were: FASTING: 40% and FED: 31%. The 95% limits for repeatability were FASTING: 29–340%, FED: 41–240%. Test reliabilities were FASTING: 0.70 and FED: 0.67. For dichotomous interpretation similar results (kappa = 0.7) were obtained using cut‐offs of [Insulin] >60 μiu/ml at T60 or T90 for FASTING and [Insulin] >51 μiu/ml at T30 or T60 for FED samples. Main limitations Oral sugar tests were performed on a small number of animals on one pasture during one season (spring). Conclusions Clinicians should beware of interpreting changes in absolute OST results owing to poor repeatability. When stabling is unavailable, OSTs of ponies at pasture may yield similar dichotomous results without prior fasting.

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Section: Accepted Articlesupporting

confidence: 68%

Section: Accepted Articlementioning

confidence: 99%