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Licensed Unlicensed Requires Authentication Published by De Gruyter July 15, 2022

Two-site evaluation of the Roche Elecsys Vitamin D total III assay

  • Julien Favresse EMAIL logo , Johan Schiettecatte , Fleur Wolff , Frederic Cotton , Marc Elsen , Christine Eucher , Catherine Rosseels and Ellen Anckaert



The high request for vitamin D testing in the last decades has led manufacturers to develop assays on automated immunoassay platforms. The objective of this study was to evaluate the performance of the new Elecsys Vitamin D total III assay for the measurement of total 25(OH)D.


A total of 844 serum samples collected in two clinical laboratories were used to evaluate the new Roche Elecsys Vitamin D total III assay. Comparisons with Roche Elecsys Vitamin D total II and liquid chromatography tandem mass spectrometry (LC-MS/MS) were carried out. Additionally, assay imprecision, linearity, matrix effects, biotin interference, cross-reactivity with 24,25(OH)2D3 and 3-epi-25(OH)D3, and outlier rate were evaluated for the Elecsys Vitamin D total III assay.


Only the comparison between LC-MS/MS and Roche Elecsys Vitamin D total III achieved the optimal specification for bias (i.e., <3.4%). Imprecision, linearity and matrix effects showed acceptable results. The biotin interference threshold was increased up to 1,200 ng/mL and the outlier rate was low (0.26%). The cross-reactivity with 24,25(OH)2D3 and 3-epi-25(OH)D3 was weak or modest in available patient samples. However, using SRM972a with a high level of 3-epi-25(OH)D3 (enriched) revealed an important cross-reactivity with both Roche Elecsys Vitamin D total II and III assays (+74.7% and +73.7%).


In conclusion, the Roche Elecsys Vitamin D total III assay presents several advantages compared to the previous assay generation: higher biotin interference threshold, broader measuring range, and better comparability with LC-MS/MS. However, the cross-reactivity toward 3-epi-25(OH)D3 is still problematic in high titer samples.

Corresponding author: Julien Favresse, Department of Laboratory Medicine, Clinique Saint-Luc Bouge, Namur, Belgium; and Department of Pharmacy, Namur Research Institute for Life Sciences, University of Namur, Namur, 5004 Bouge, Belgium, Phone: +32 81 20 91 44, E-mail:

  1. Research funding: None declared.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: Authors state no conflict of interest.

  4. Informed consent: Informed consent is not required for the evaluation of a new generation of assay.

  5. Ethical approval: The local Institutional Review Board deemed the study exempt from review.


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Supplementary Material

The online version of this article offers supplementary material (

Received: 2022-02-25
Accepted: 2022-07-03
Published Online: 2022-07-15
Published in Print: 2022-09-27

© 2022 Walter de Gruyter GmbH, Berlin/Boston

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