25-OH-Vitamin D ELISA

Intended Purpose

Enzyme immunoassay for the quantitative measurement of total 25-OH-Vitamin D (25-OH-Vitamin D2 and 25-OH-Vitamin D3) in human serum as a physiological marker as an aid to the diagnosis of vitamin D sufficiency. Patient population includes individuals who are at risk to deviate from sufficient levels of vitamin D. The 25-OH-Vitamin D ELISA is a solid phase enzyme-linked immunosorbent assay (ELISA), based on the principle of competitive binding and measured on an absorbance reader. The assay is semi-automated requiring general purpose laboratory instruments and consumables such as absorbance microplate reader/washer, vortexer and pipettes to execute the test. The assay is adaptable by laboratory personnel to automate on open ELISA based liquid handler platforms; however, the programming of the steps and timing required by the manual kit assay test instructions must be strictly adhered to and verified by the laboratory. Test results are calculated from a standard curve and compared to defined cut-off. The test kit is intended for professional laboratory use by trained personnel. The test kit is not for home or lay person use.

Clinical relevance

The accurate assessment of a person’s vitamin D status relies on the measurement of 25-OH-vitamin D total (25-OH-vitamin D2 + 25-OH-vitamin D3) levels. Therefore, our assay has been developed to detect both D2 and D3 variants. Balanced levels of vitamins in the human body are important to prevent disease (e.g. cardiovascular diseases, type 2 diabetes)^[3]. Among essential vitamins, vitamin D is important to maintain calcium metabolism and bone health^[2]. Additionally, vitamin D has received interest for its role in non-skeletal diseases and disorders^[4,5]. It has been found that a large fraction of patients encountered in clinical practice are deficient in vitamin D^[1]. Identifying patients with insufficient vitamin D levels in patient populations at risk from hypovitaminosis D, can help to maintain health and prevent disease^[2].

Correlation to LC-MS/MS

The Tecan-IBL 25-OH-Vitamin D ELISA is correlated against the IDHPLC-MS/MS reference method (The method is recognized by the Joint Committee for Traceability in Laboratory Medicine (JCTLM) as a reference measurement procedure (RMPs) of a higher-order).

The correlation was performed on 40 serum samples (CDC phase I samples, single-donor sera) comparing the results of TECAN-IBL 25-OH-Vitamin D ELISA to reference concentrations assigned by CDC using IDHPLC-MS/MS. The regression analysis showed a slope of 0.9787, an intercept of -2.961 nmol/L (nmol/L = 2.5 x ng/mL) with a correlation of r=0.94.

Automation

The Tecan-IBL 25-OH-Vitamin D ELISA has been developed to be adaptable to open ELISA processors. An in-house correlation has been performed comparing the manual work-up of the 25-OH-Vitamin D ELISA and the automated work-up of the 25-OH-Vitamin D ELISA on the TECAN Freedom EVOlyzer®.

Advantages of our ELISA

• Excellent correlation to the IDHPLC-MS/MS reference method RE53041 = 0.9787 x LCMS – 2.961, R = 0.94
• Direct assay without extraction
• Ready-to-use reagents
• Easily adaptable on open ELISA platforms

*Product availability and regulatory status may vary across regions outside the EU depending on local country-specific registration. Consult with your Tecan associate for further information.

For concrete data please consult the Instruction for Use in the download box on the top right side.