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Clinical Chemistry 53: 1104-1108, 2007. First published April 13, 2007; 10.1373/clinchem.2006.077560
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Right arrow Endocrinology and Metabolism
(Clinical Chemistry. 2007;53:1104-1108.)
© 2007 American Association for Clinical Chemistry, Inc.

Endocrinology and Metabolism

Evaluation of a 1,25-Dihydroxyvitamin D Enzyme Immunoassay

Isolde Seiden-Long1 and Reinhold Vieth1,2,a

1 Department of Laboratory Medicine and Pathobiology, University of Toronto Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Canada.
2 Department of Nutritional Sciences, University of Toronto, Toronto, Canada.

aAddress correspondence to this author at: Pathology and Laboratory Medicine, Mount Sinai Hospital, 600 University Ave., Toronto, Ontario, M5G 1X5 Canada. Fax 416-586-8628; e-mail rvieth{at}mtsinai.on.ca.

Background: Radioactive reagents are used in most assays for measurement of 1,25-dihydroxyvitamin D [1,25(OH)2D]. We evaluated a 1,25(OH)2D enzyme immunoassay (EIA) from IDS Ltd. that uses solid-phase immunoextraction and colorimetric detection and compared results to those of the thymus radioreceptor assay (RRA) for 1,25(OH)2D.

Methods: We collected serum samples (n = 145) representing an even distribution (0–200 pmol/L) of 1,25(OH)2D concentrations and Vitamin D External Quality Assessment Scheme (DEQAS) proficiency survey samples from 2004 surveys (n = 15) and stored them at –20 °C. We analyzed all samples with both EIA and RRA methods. We calculated imprecision using 5 QC samples in quadruplicate in each run (n = 6), including both pooled patient material used for QC with the RRA and QC material included in the EIA reagent set. We evaluated calibration stability by analyzing calibrators from different lots on the same plate and determining if calculated sample values drifted significantly.

Results: Deming linear regression between IDS EIA and RRA methods yielded slope 1.25 (95% CI 1.13–1.37), y-intercept –3 (95% CI –18 to 12), R2 = 0.74. DEQAS proficiency survey samples for 2004 were all within 30% of the all-methods-trimmed mean. Imprecision CVs were 12%–16% within-run and 15%–20% between-run.

Conclusions: We find no evidence of inferiority to the classic calf-thymus receptor assay for 1,25(OH)2D and no disadvantage in the results generated by the IDS EIA using samples from the major proficiency survey for 1,25(OH)2D. According to the product insert, however, the IDS EIA underestimates 1,25(OH)2D2 compared with the D3 form.






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Copyright © 2007 by the American Association for Clinical Chemistry.