Test Catalog

Test Id : 2425D

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25-Hydroxyvitamin D:24,25-Dihydroxyvitamin D Ratio, Serum

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Overview

Test Catalog

Useful For
Suggests clinical disorders or settings where the test may be helpful

As a screening test for inactivating CYP24A1 mutations in patients with symptoms, signs, or biochemical findings of parathyroid hormone (PTH)-independent hypercalcemia or hypercalciuria

Profile Information
A profile is a group of laboratory tests that are ordered and performed together under a single Mayo Test ID. Profile information lists the test performed, inclusive of the test fee, when a profile is ordered and includes reporting names and individual availability.

Test Id Reporting Name Available Separately Always Performed
2425R 24,25 Dihydroxy Vitamin D No Yes
25HDN 25-Hydroxyvitamin D2 and D3, S Yes Yes

Method Name
A short description of the method used to perform the test

Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

Portions of this test are covered by patent(s) held by Quest Diagnostics

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.

Yes

Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

25HDN:24,25 Dihydroxy VitD Ratio, S

Aliases
Lists additional common names for a test, as an aid in searching

2425D

2425R

2425

24,25-Dihydroxyvitamin D

Hydroxycalcifediol

Hydroxycalcidiol

P450CC24

Cytochrome P450 Family 24

Cytochrome P450 Subfamily XXIV

Vitamin D 24-Hydroxylase

25-Hydroxyvitamin D 24-Hydroxylase

Dihydrocholesterol

2425OHD

CYP24A1

Calciferol

Ergocalciferol

Calcidiol

Cholecalciferol

Colecalciferol

Vitamin D Assay

CYP24

Specimen Type
Describes the specimen type validated for testing

Serum

Ordering Guidance

Please consider the 25HDN or DHVD tests for Vitamin D assessment. The 25-hydroxyvitamin D test (25HDN / 25-Hydroxyvitamin D2 and D3, Serum) in serum is the preferred initial test for assessing vitamin D status and most accurately reflects the body's vitamin D stores. In the presence of renal disease or hypercalcemia, testing of 1,25-dihydroxy vitamin D (DHVD) might be needed to adequately assess vitamin D status.

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Container/Tube:

Preferred: Red top

Specimen Volume: 3 mL

Collection Instructions: Spin down within 2 hours of draw.

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the Testing Laboratory

1.1 mL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

Gross hemolysis OK
Gross lipemia Reject
Gross icterus OK

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Serum Refrigerated (preferred) 14 days
Frozen 30 days
Ambient 7 days

Useful For
Suggests clinical disorders or settings where the test may be helpful

As a screening test for inactivating CYP24A1 mutations in patients with symptoms, signs, or biochemical findings of parathyroid hormone (PTH)-independent hypercalcemia or hypercalciuria

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

Vitamin D is a generic designation for a group of fat-soluble, structurally similar sterols. The 25HDN / 25-Hydroxyvitamin D2 and D3, Serum assay is the preferred initial test for assessing vitamin D status and most accurately reflects the body's vitamin D stores. In the presence of renal disease, DHVD / 1,25-Dihydroxyvitamin D, Serum testing might be needed to adequately assess vitamin D status. For patients with loss of function inactivating CYP24A1 mutations, this test (2425D / 25-Hydroxyvitamin D2 and D3:24,25-Dihydroxyvitamin D Ratio, Serum) may be helpful

 

Loss of function mutations in the CYP24A1 gene have been shown to lead to insufficient deactivation of bioactive vitamin D metabolites, resulting in a phenotype characterized by suppressed serum parathyroid hormone (PTH), increased serum 1,25-dihydroxyvitamin D (DHVD) concentrations, hypercalcemia, and hypercalciuria or nephrolithiasis.

 

Vitamin D compounds in the body are exogenously derived by dietary means; from plants as 25-hydroxyvitamin D2 (ergocalciferol or calciferol) or from animal products as 25-hydroxyvitamin D3 (cholecalciferol or calcidiol). Vitamin D may also be endogenously derived by conversion of 7-dihydrocholesterol to 25-hydroxyvitamin D3 in the skin upon ultraviolet exposure.

 

25-Hydroxyvitamin D (25HDN) is subsequently formed by hydroxylation (CYP2R1) in the liver. 25HDN is a prohormone that represents the main reservoir and transport form of vitamin D, being stored in adipose tissue and tightly bound by a transport protein while in circulation. Biological activity is expressed in the form of DHVD the active metabolite of 25HDN. 1-Alpha-hydroxylation (CYP27B1) occurs on demand, primarily in the kidneys, under the control of parathyroid hormone (PTH) before expressing biological activity. Like other steroid hormones, DHVD binds to a nuclear receptor, influencing gene transcription patterns in target organs.

 

25HDN may also be converted into the inactive metabolite 24,25-dihydroxyvitamin D (24,25D) by (CYP24A1) hydroxylation. This process regulated by parathyroid hormone (PTH) might increase DHVD synthesis at the expense of the alternative hydroxylation (CYP24A1) product 24,25D. Inactivation of 25HDN and DHVD by CYP24A1 is a crucial process that prevents over production of DHVD and resultant vitamin D toxicity.

 

DHVD stimulates calcium absorption in the intestine and its production is tightly regulated through concentrations of serum calcium, phosphorus, and PTH. DHVD promotes intestinal calcium absorption and, in concert with PTH, skeletal calcium deposition, or less commonly, calcium mobilization. Renal calcium and phosphate reabsorption are also promoted, while prepro-PTH mRNA expression in the PTH glands is downregulated. The net result is a positive calcium balance, increasing serum calcium and phosphate levels, and falling PTH concentrations. In addition to its effects on calcium and bone metabolism, DHVD regulates the expression of a multitude of genes in many other tissues including immune cells, muscle, vasculature, and reproductive organs.

 

DHVD levels are decreased in hypoparathyroidism and in chronic renal failure. DVHD levels may be high in primary hyperparathyroidism and in physiologic hyperparathyroidism secondary to low calcium or vitamin D intake. Some patients with granulomatous diseases (eg, sarcoidosis) and malignancies containing nonregulated 1-alpha hydroxylase in the lesion might have hypercalcemia that appears vitamin D mediated with normal or high serum phosphate (hyperphosphatemia) and hypercalcemia (both of which might be severe), in addition to low parathyroid hormone (PTH) and absent parathyroid hormone-related peptide (PTHRP). Differential diagnostic considerations include vitamin D intoxication and CYP24A1 deficiency

Reference Values
Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.

Interpretative commentary provided based on 25-hydroxyvitamin D (25HDN) to 24,25-dihydroxyvitamin D (24,25D) ratio result.

 

25HDN to 24,25D ratio less than 25*

*Interpretation: Normal (Ratio of less than 25 may also be observed in heterozygous carriers of CYP24A1 mutations)

 

25HDN to 24,25D ratio between 25-80**

**Interpretation: Ratios in the 25 to 80 range can be seen in patients with low vitamin D or heterozygous CYP24A1 mutations. Confirmation with molecular testing is recommended.

 

25HDN to 24,25D ratio greater than 80***

***Interpretation: Ratios greater than 80 indicate probable biallelic CYP24A1 mutation or deletion. Confirmation with molecular testing is recommended.

 

Reference values not applicable for 24,25 Dihydroxyvitamin D Total result.

 

Results should be interpreted in the context of other biochemical findings including serum calcium, parathyroid hormone, and 1,25 dihydroxyvitamin D concentrations. If 25-OH-D is less than 20 ng/ml the ratio of 25-OH-D to 24,25-dihydroxyvitamin D will be falsely elevated since there is no inactivation 25-OH-D to 24,25-dihydroxyvitamin D.

Interpretation
Provides information to assist in interpretation of the test results

Results should be interpreted in the context of other biochemical findings including serum calcium, parathyroid hormone (PTH), and 1,25 dihydroxyvitamin D (DHVD) concentrations. If 25-hydroxyvitamin D (25HDN) result is less than 20 ng/mL, the ratio of 25-OH-D to 24,25-dihydroxyvitamin D (24,25D) will be falsely elevated since there is no inactivation of 25-OH-D to 24,25D.

 

24,25D formation by CYP24A1 is dependent on CYP24A1 activity and the concentrations of its substrate, 25HDN. The ratio of 25HDN to 24,25D, therefore, allows the most reliable estimation of CYP24A1 activity.

 

Ratios of 25HDN to 24.25D less than 25 may be interpreted as normal, though ratio of less than 25 may also be observed in heterozygous carriers of CYP24A1 mutations.

 

Ratios of 25HDN to 24.25D between the 25 and 80 range may be seen in patients with low vitamin D or heterozygous CYP24A1 mutations. Confirmation with molecular testing is recommended.

 

Confirmation with molecular testing is also recommended for ratios of 25HDN to 24.25D greater than 80, as this may indicate a probable biallelic CYP24A1 mutation or deletion.

Cautions
Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

Because of the substrate dependency of the 25-hydroxyvitamin D (25HDN) to 24,25-dihydroxyvitamin D (24,25D)  ratio, it is essential for accurate determination of this ratio that 25HDN to 24,25D are measured in the same draw and sample using the same methodologies for 25HDN to 24,25D that were used when the ratio reference ranges were established. This is an important consideration for clinicians and clinical chemists who recommend this testing, because the absolute value of serum 25HDN to 24,25D can be misleading if calculated from the 2 separate measurements.

 

False-low 25HDN to 24,25D ratios could lead to delayed diagnosis of CYP24A1 deficiency. False-high ratios might cause unnecessary molecular testing.

Clinical Reference
Recommendations for in-depth reading of a clinical nature

1. Kaufmann M, Gallagher JC, Peacock M, et al: Clinical utility of simultaneous quantitation of 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D by LC-MS/MS involving derivatization with DMEQ-TAD. J Clin Endocrinol Metab 2014;July 99(7):2567-2574. doi: 10.1210/jc.2013-4388

2. Ketha H, Kumar R, Singh RJ: LC-MS/MS for Identifying Patients with CYP24A1 Mutations. Clin Chem 2016 Jan;62(1):236-242

Method Description
Describes how the test is performed and provides a method-specific reference

Analytes of interest and deuterated internal standard are extracted, derivatized and analyzed by liquid chromatography-tandem mass spectrometry.(Unpublished Mayo method)

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information

No

Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Tuesday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

2 to 6 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

2 weeks

Performing Laboratory Location
Indicates the location of the laboratory that performs the test

Rochester

Fees
Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

  • Authorized users can sign in to Test Prices for detailed fee information.
  • Clients without access to Test Prices can contact Customer Service 24 hours a day, seven days a week.
  • Prospective clients should contact their Regional Manager. For assistance, contact Customer Service.

Test Classification
Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

This test was developed, and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information
Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.

82306

82542

LOINC® Information

Test Id Test Order Name Order LOINC Value
2425D 25HDN:24,25 Dihydroxy VitD Ratio, S 94674-9
Result Id Test Result Name Result LOINC Value
Result LOINC Value Tooltip

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Pdf

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports