25OH Vitamin D Total Assay - University of California, San Francisco

I. PURPOSE
UNIVERSITY OF CALIFORNIA SAN FRANCISCO, CLINICAL LABORATORIES
The LIAISON 25 OH Vitamin D TOTAL Assay uses chemiluminescent immunoassay (CLIA)
technology for the quantitative determination of 25-hydroxyvitiamin D and other hydroxylated vitamin
D metabolites in human serum to be used in the assessment of vitamin D sufficiency. Assay results
should be used in conjunction with other clinical or laboratory data to assist the clinician in making
individual patient management decisions in an adult population.
SUMMARY AND EXPLANATION OF TEST
The role of vitamin D in bone and mineral metabolism was recognized from its first identification as a
factor that could cure rickets. However, vitamin D is now recognized as a prohormone, which has
multiple roles in maintaining optimal health.
Vitamin D3 (cholecalciferol) and vitamin D2 (ergocalciferol) are the most abundant forms of vitamin D
in the body. Vitamin D is synthesized in the skin from 7-dehydrocholesterol in response to sunlight.
The best nutritional sources of D3 are oily fish, primarily salmon and mackerel. Vitamin D2‘s nutritional
sources are some vegetables, yeast and fungi. The vegetarian diet is abundant in vitamin D2.
Vitamin D (D3, D2 and metabolites) is converted to 25-hydroxyvitamin D in the liver. The
measurement of 25-OH vitamin D concentration in the serum or plasma is the best indicator of
vitamin D nutritional status.
The optimal level of 25-OH vitamin D is subject to some debate, but >32ng/mL (>80nmol/L) is
accepted as sufficient for bone health. Vitamin D toxicity is a recognized problem, but a rare
occurrence. Instead, a recent growing public health problem is vitamin D insufficiency. Several
research studies have identified widespread vitamin D insufficiency in apparent healthy populations
world-wide.
Vitamin D deficiency is commonly treated with vitamin D2 or D3 medications in doses ranging from
50,000 IU/month to 50,000 IU/week. Fortified foods and nutrition supplements may contain either
form. To ensure accurate assessment of the total vitamin D all forms, including D3, D2 and
metabolites, must be measured.
Recent studies have identified an inactive 3-epimer of 25-OH vitamin D, which may be present in the
serum of infants under the age of one year. Thus it is important that an assay avoid measuring the
inactive 3-epi form and only measure the active D3 and D2 forms equally.
BIOLOGICAL PRINCIPLES OF THE PROCEDURE
The LIAISON 25 OH Vitamin D assay is a direct competitive chemiluminescence immunoassay
(CLIA) for quantitative determination of total 25 OH vitamin D in serum. During the first incubation, 25
OH vitamin D is dissociated from its binding protein and binds to the specific antibody on the solid
phase. After 10 minutes, the tracer (vitamin D linked to an isoluminol derivative) is added. After a
second 10 minute incubation, the unbound material is removed with a wash cycle. Subsequently, the
starter reagents are added to initiate a flash chemiluminescent reaction. The light signal is measured
by a photomultiplier as relative light units (RLU) and is inversely proportional to the concentration of
25 OH vitamin D present in calibrators, controls or samples.
II. POLICY/SCOPE
This procedure is intended for use by licensed CLS staff in the China Basin Chemistry section.
III. TEST AVAILABILITY
Test performed Monday, Wednesday and Fridays. Results will be available by 16:00.
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