Comparison of 11 human insulin assays: implications for clinical investigation and research

Clin Chem. 2007 May;53(5):922-32. doi: 10.1373/clinchem.2006.077784. Epub 2007 Mar 15.

Abstract

Background: The American Diabetes Association task force on standardization of insulin assays in 1996 showed wide variation in assay bias. Newer assays are specific for insulin, with several now available on automated immunoassay analyzers.

Methods: In 2004, we compared 11 commercially available insulin assays by analyzing 150 serum samples (99 fasting/51 postprandial) from study participants with various degrees of glucose intolerance (exclusions being type 1 diabetes, insulin treatment, or presence of insulin antibodies). All assays were calibrated against International Reference Preparation 66/304. One assay was not specific for insulin and another was an RIA; 10 assays used enzyme/chemiluminescent labels. Bland-Altman difference plots were modified to use the mean insulin from all assays on the x-axis as a common comparator.

Results: As in the 1996 study, insulin values from the different assays varied by a factor of 2, with the nonspecific assay ranking in the middle of the distribution. Spearman rank correlation coefficients, for ranking samples vs the mean, were 0.983-0.997. Both offsets and concentration-dependent differences were seen in the modified difference plots. Imprecision (mean CV) for automated assays (3%) was not significantly different from manual assays (5%). Similar values were obtained when one automated assay was run in laboratories in both the UK and the US. Results of 1 assay showed lower insulin concentrations in heparinized plasma than in serum.

Conclusions: Assay performance must be considered before comparing insulin results. The 2-fold variation in insulin results may be related to specificity, manufacturers' calibration procedures or conversion factors.

Publication types

  • Comparative Study
  • Multicenter Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Anticoagulants
  • Autoanalysis
  • Glucose Intolerance / blood
  • Heparin
  • Humans
  • Immunoassay / methods*
  • Immunoassay / standards
  • Insulin / blood*
  • Linear Models
  • Middle Aged
  • Plasma
  • Proinsulin / blood
  • Serum

Substances

  • Anticoagulants
  • Insulin
  • Heparin
  • Proinsulin