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1 Departments of Pediatrics, Biochemistry and Molecular Biology, Medical University of South Carolina, 171 Ashley Ave., Charleston, SC 29425. Fax 843-792-1844; e-mail Hollisb{at}musc.edu
Background: Measurement of circulating 25-hydroxyvitamin D [25(OH)D] is important in the management of metabolic bone disease. The aim of this study was to compare two widely used methods for the quantification of circulating 25(OH)D with attention to their abilities to measure 25-hydroxylated ergocalciferol (vitamin D2) [25(OH)D2] and cholecalciferol (vitamin D3) [25(OH)D3].
Methods: We used two commercially available, Food and Drug Administration-approved, radioiodine (125I)-based RIA kits for the detection of 25(OH)D (DiaSorin, Stillwater, MN and IDS Ltd, Tyne and Wear, United Kingdom). These methods were tested for general assay performance, including antibody specificity. Results were compared with those of an HPLC-based direct ultraviolet detection method.
Results: Within- and between-run CVs were 
10%. Both methods
quantitatively recovered 25(OH)D3 added to serum, but only
the DiaSorin kit quantitatively recovered 25(OH)D2. The
primary antibody in the IDS kit had unequal reactivities with pure
25(OH)D2 and 25(OH)D3, whereas the DiaSorin
primary antibody reacted with them equally. In 50 patient samples
assayed by HPLC, the IDS method, but not the DiaSorin method,
underestimated total circulating 25(OH)D when significant circulating
25(OH)D2 was present in patient samples.
Conclusions: Some immunoassays may underestimate total 25(OH)D when 25(OH)D2 constitutes an appreciable part of the total.
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S. Levis, A. Gomez, C. Jimenez, L. Veras, F. Ma, S. Lai, B. Hollis, and B. A. Roos Vitamin D Deficiency and Seasonal Variation in an Adult South Florida Population J. Clin. Endocrinol. Metab., March 1, 2005; 90(3): 1557 - 1562. [Abstract] [Full Text] [PDF]
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J.-C. Souberbielle, V. Fayol, C. Sault, E. Lawson-Body, A. Kahan, and C. Cormier Assay-Specific Decision Limits for Two New Automated Parathyroid Hormone and 25-Hydroxyvitamin D Assays Clin. Chem., February 1, 2005; 51(2): 395 - 400. [Abstract] [Full Text] [PDF]
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J.-C. Souberbielle, E. Lawson-Body, B. Hammadi, E. Sarfati, A. Kahan, and C. Cormier The Use in Clinical Practice of Parathyroid Hormone Normative Values Established in Vitamin D-Sufficient Subjects J. Clin. Endocrinol. Metab., August 1, 2003; 88(8): 3501 - 3504. [Abstract] [Full Text] [PDF]
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J.-C. Souberbielle, E. Lawson-Body, and C. Kindermans Authors' Response: Vitamin D Status and Redefining Serum PTH Reference Range in the Elderly J. Clin. Endocrinol. Metab., February 1, 2002; 87(2): 947 - 947. [Full Text] [PDF]
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