A Comparison of Two Immunoassays for Analysing Plasma 25- hydroxyvitamin D

Kurtis Sarafin, Nicolas Hidiroglou, Stephen P.J. Brooks*
Nutrition Research Division, E315-PL2203E Banting Research Centre, 251 Sir Frederick Banting Driveway, Ottawa, ON K1A 0K9, Canada.

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© 2011 Sarafin et al;

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Nutrition Research Division, E315-PL2203E Banting Research Centre, 251 Sir Frederick Banting Driveway,Ottawa, ON K1A 0K9, Canada; Tel: 613-957-0932; Fax: 613-941-0451; E-mail:


A total of 1628 human plasma samples from Cycle 1 of the Canadian Health Measures Survey were assayed for total 25-hydroxyvitamin D using the DiaSorin RIA method and the Diasorin ”LIAISON 25 OH Vitamin D Total“ method. Bland-Altman comparison showed an average bias of 4.8 ± 16.7 nmol/L (6.3%: P<0.001) with the LIAISON method giving higher values. The relationship was investigated using linear and Deming regression. Linear regression gave: LIAISON = RIA±(0.87 ± 0.02) + (13.3 ± 1.2) (mean ± SE) and weighted Deming regression (constant CV) gave: LIAISON = RIA±(1.14 ± 0.02) - (4.2 ± 1.2). The significant deviations from a slope of unity and the significant non-zero intercepts were further investigated using non-linear regression. Quadratic regression gave: LIAISON = RIA2 ± (-0.0025 ± 0.0005) + RIA±(1.211 ± 0.07) + (2.9 ± 2.5). The intercept was not significantly different from 0. The quadratic equation significantly decreased the residual sum of squares (P<0.0001) indicating this model better described the relationship. Non-linearity was apparent at RIA 25-hydroxyvitamin D ≥ 110 nmol/L, where the relationship was described by: LIAISON = 97.5339 + 0.1388±RIA (r2 = 0.0039; N.S.). However, removing points RIA ≥ 110 nmol/L did not substantially alter the regression parameters. Comparing the analytical imprecision with the total random regression error (Sy/x) suggested that sample-related effects were not present. It is recommended that cross-over analysis between these two methods include points from all parts of the range of interest to elucidate the complete nature of the relationship.

Keywords: Vitamin D, Liaison, RIA, regression, comparison.