What is the correct laboratory test for ordering Vitamin D (25-hydroxyvitamin D) levels?

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Last updated: April 3, 2025View editorial policy

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From the Guidelines

The correct lab test for ordering vitamin D is the 25-hydroxyvitamin D (25(OH)D) blood test. This test measures the total circulating vitamin D in the body and is the most accurate indicator of vitamin D status, as stated in the study by 1. When ordering this test, healthcare providers should specify "25-hydroxyvitamin D" or "25(OH)D" rather than simply "vitamin D" to ensure the correct test is performed.

Key Considerations

  • The normal range for 25(OH)D levels is typically considered to be 30-100 ng/mL (75-250 nmol/L), with levels below 20 ng/mL indicating deficiency, as noted in the study by 1.
  • This test should be ordered for patients with risk factors for vitamin D deficiency, including those with limited sun exposure, darker skin, obesity, malabsorption disorders, or certain medications that affect vitamin D metabolism.
  • The 1,25-dihydroxyvitamin D test (the active form) is not recommended for routine vitamin D screening as it does not accurately reflect vitamin D stores and is primarily used for specific conditions like kidney disease or certain metabolic disorders, as mentioned in the study by 1.
  • Testing is best performed after fasting, though not strictly required, and levels may vary seasonally with higher values typically seen in summer months.

Laboratory Measurement

The study by 1 highlights the importance of choosing an assay that measures both 25(OH)D2 and 25(OH)D3, and participating in an external quality control scheme to ensure accurate results. The study by 1 also notes the limited effects of standardization programs on improving the accuracy of immunoassay methods, emphasizing the need for careful selection of laboratory tests.

Clinical Implications

Healthcare providers should prioritize the 25(OH)D test for assessing vitamin D status, given its accuracy and relevance to clinical decision-making, as supported by the studies by 1. By doing so, they can ensure that patients receive appropriate diagnosis and treatment for vitamin D deficiency, ultimately improving morbidity, mortality, and quality of life outcomes.

From the Research

Correct Lab for Ordering Vitamin D

To determine the correct lab for ordering vitamin D, it is essential to consider the accuracy and reliability of different assay methods. The following points highlight the key findings from various studies:

  • The measurement of 25-hydroxyvitamin D (25[OH]D) is considered the best marker of vitamin D status 2.
  • Automated immunoassays and tandem mass spectrometry are the most widely used assays for the measurement of 25(OH)D, but significant differences remain among various methods and laboratories 2.
  • Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a reliable and robust method for the measurement of 25(OH)D in human serum, with high accuracy and precision 3, 4, 5.
  • Comparison of LC-MS/MS with radioimmunoassay (RIA) and automated immunoassays showed significant differences in results, highlighting the need for standardization and harmonization of 25(OH)D measurements 3, 4, 5, 6.
  • The use of well-standardized assays in clinical laboratories is crucial to ensure accurate and reliable results for vitamin D testing 6.

Key Considerations for Vitamin D Testing

When ordering vitamin D tests, consider the following:

  • The choice of assay method: LC-MS/MS is a reliable and accurate method, but other methods like RIA and automated immunoassays may also be used.
  • The laboratory's experience and expertise in vitamin D testing.
  • The need for standardization and harmonization of 25(OH)D measurements to ensure accurate and reliable results.
  • The potential for significant differences in results between various assays and laboratories, highlighting the importance of using well-standardized assays 6.

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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