Assay Variation Confounds the Diagnosis of Hypovitaminosis D: A Call for Standardization
N. Binkley,
D. Krueger,
C. S. Cowgill,
L. Plum,
E. Lake,
K. E. Hansen,
H. F. DeLuca and
M. K. Drezner
Department of Medicine Osteoporosis Clinical Center and Research Program (N.B., D.K., C.S.C., K.E.H., M.K.D.); Institute on Aging (N.B., D.K., C.S.C., K.E.H., M.K.D.); Geriatric Research Education and Clinical Center, William S. Middleton Veterans Affairs Medical Center (M.K.D.); and Department of Biochemistry (L.P., E.L., H.F.D.), University of Wisconsin-Madison, Madison, Wisconsin 53705
Address all correspondence and requests for reprints to: Neil Binkley, M.D., 2870 University Avenue, Suite 100, Madison, Wisconsin 53705. E-mail: nbinkley{at}facstaff.wisc.edu.
Endemic hypovitaminosis D contributes to osteoporosis development.However, variation in 25-hydroxyvitamin D (25OHD) measurementis reported and confounds the diagnosis of vitamin D insufficiency/deficiency.This report emphasizes the marked variability observed in serum25OHD measurements between laboratories.
Initially, postmenopausal women had serum 25OHD determinations:42 in laboratory A, 20 in laboratory B. Their mean (SEM) serum25OHD concentrations were 46 (2.1) and 21 (2.3) ng/ml in laboratoriesA and B, respectively. Furthermore, there was little overlapin serum 25OHD among these clinically similar individuals. Specifically,17% of those measured in laboratory A but 90% in laboratoryB were below an arbitrary threshold value of 32 ng/ml.
Subsequently, serum was obtained from 10 healthy adults. Twoaliquots from each individual, one of which was spiked with20 ng/ml 25OHD, were sent to six laboratories. Substantial variabilitywas noted between these six laboratories. The mean serum 25OHDconcentration ranged from 17.1–35.6 ng/ml. Similarly,the mean increase produced by spiking with 20 ng/ml ranged from7.7–18.0 ng/ml.
In conclusion, 25OHD assays yield markedly differing results;whether an individual is found to have low or normal vitaminD status is a function of the laboratory used. If the medicalcommunity is to make progress in correcting widespread hypovitaminosisD, 25OHD measurement must be standardized.
Abbreviation: 25OHD, 25-Hydroxyvitamin D.
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162(6):
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October 1, 2006;
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[Full Text][PDF]
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90(11):
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August 1, 2005;
90(8):
4566 - 4572.
[Abstract][Full Text][PDF]
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J. Clin. Endocrinol. Metab.,
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90(6):
3800 - 3801.
[Full Text][PDF]
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90(6):
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June 1, 2005;
51(6):
1072 - 1074.
[Full Text][PDF]
P. Glendenning and W. D. Fraser 25-OH-Vitamin D Assays
J. Clin. Endocrinol. Metab.,
May 1, 2005;
90(5):
3129 - 3129.
[Full Text][PDF]
M. F. Holick 25-OH-Vitamin D Assays
J. Clin. Endocrinol. Metab.,
May 1, 2005;
90(5):
3128 - 3129.
[Full Text][PDF]
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J. Clin. Endocrinol. Metab.,
March 1, 2005;
90(3):
1557 - 1562.
[Abstract][Full Text][PDF]
B. W. Hollis, C. L. Wagner, A. Kratz, P. M. Sluss, and K. B. Lewandrowski Normal Serum Vitamin D Levels
N. Engl. J. Med.,
February 3, 2005;
352(5):
515 - 516.
[Full Text][PDF]
D. A. Hanley and K. S. Davison Vitamin D Insufficiency in North America
J. Nutr.,
February 1, 2005;
135(2):
332 - 337.
[Abstract][Full Text][PDF]
E. Seeman and G. J. Strewler Clinical and Basic Research Papers - November 2004 Selections
IBMS BoneKEy,
December 1, 2004;
1(12):
1 - 2.
[Full Text][PDF]
R. P Heaney Functional indices of vitamin D status and ramifications of vitamin D deficiency
Am. J. Clinical Nutrition,
December 1, 2004;
80(6):
1706S - 1709S.
[Abstract][Full Text][PDF]
G. D. Carter, R. Carter, J. Jones, and J. Berry How Accurate Are Assays for 25-Hydroxyvitamin D? Data from the International Vitamin D External Quality Assessment Scheme
Clin. Chem.,
November 1, 2004;
50(11):
2195 - 2197.
[Full Text][PDF]
Measuring Serum Vitamin D Is Problematic
Journal Watch Gastroenterology,
August 31, 2004;
2004(831):
10 - 10.
[Full Text]
Measuring Serum Vitamin D is Problematic
Journal Watch (General),
August 13, 2004;
2004(813):
2 - 2.
[Full Text]
E. Seeman and G. J. Strewler Clinical and Basic Research Papers - May and June 2004 Selections
IBMS BoneKEy,
July 1, 2004;
1(7):
1 - 3.
[Full Text][PDF]
B. W. Hollis Editorial: The Determination of Circulating 25-Hydroxyvitamin D: No Easy Task
J. Clin. Endocrinol. Metab.,
July 1, 2004;
89(7):
3149 - 3151.
[Full Text][PDF]
