The determination of detection limits for insulin antibody assays.
In order to define the detection limit of a radioimmunoassay for insulin-antibody a correction was made for binding in the presence of an excess of unlabelled insulin and assay precision was calculated. One hundred forty control sera were assessed; all were islet cell antibody negative. For each sample, binding of 125-I human insulin was determined both with and without excess unlabelled insulin, subtraction of the latter acting as a correction. The distribution of uncorrected binding was skewed while corrected binding was normally distributed, (mean (SD) = 0.149 (0.298%)) Precision, defined as the mean of the standard deviations of replicates, was 0.263%. Detection limits calculated from the estimate of precision (0.263%) or from the standard deviation of the corrected binding (0.298%) were similar. Two hundred thirty sera from insulin-treated patients were studied. Precision was plotted as a 'precision profile' and the detection limit calculated from the precision for binding of less than 1% [0.261%]; 88% of the sera were positive [cut-off 1.3%, p less than 0.01]. We conclude that corrected binding is normally distributed in antibody-negative sera and that an estimate of assay precision can be used to define the detection limit of the assay.
Middlesex Hospital, London, UK.
A B
AB
Kurtz
L
L
DiSilvio
E
E
Bosi
D000936:Antigen-Antibody Complex / Q000032:analysis; D003922:Diabetes Mellitus, Type 1 / Q000188:drug therapy / Q000276:immunology*; D006801:Humans; D007330:Insulin Antibodies / Q000032:analysis*; D007457:Iodine Radioisotopes; D012016:Reference Values
D000936:Antigen-Antibody Complex; D007330:Insulin Antibodies; D007457:Iodine Radioisotopes