Department of Clinical Biochemistry, Aarhus University Hospital, Denmark.
Clin Chem Lab Med. 2012 Nov;50(11):1965-8. doi: 10.1515/cclm-2011-0964.
Vitamin D deficiency is considered a major health issue and therefore there is a need for reliable routine tests for measurement of the vitamin in blood samples. Here we present a validation of the recently released Roche 25-OH Vitamin D Total assay (Vitamin D Total).
We analyzed control materials (2 levels) and patient serum pools (3 levels) ranging from 34 to 123 nmol/L 84 times over a period of 21 days, and we analyzed five serum pools in 10 separate runs to verify the limit of quantification. We also analyzed 53 paired samples of serum and Li-heparin plasma. We evaluated the 25-OH Vitamin D Total assay in comparison to our in-house liquid chromatography tandem mass spectrometry (LC-MS/MS) method [194 patient samples without 25-hydroxy vitamin D(2) (25OHD(2)) and 23 patient samples containing 25OHD(2)].
At concentrations of 34 and 56 nmol/L within-run CVs were 4.8% and 1.9% and total CVs were 8.3% and 6.1%. We verified that the limit of quantification was 22.5 nmol/L, as stated by the manufacturer. No significant difference was observed between serum and plasma samples (Li-heparin). Comparison with LC-MS/MS using 194 samples containing 25OHD(3) only (no 25OHD(2)) showed Vitamin D Total nmol/L=1.07×(LC-MS/MS) nmol/L+4.7 nmol/L, whereas comparison of 25OHD(2) using 23 patient samples showed Vitamin D Total nmol/L=0.55×(LC-MS/MS) nmol/L-2.38 nmol/L (Demings regression).
The Roche Vitamin D Total assay is judged suitable for measurement of 25OHD in serum and Li-heparin plasma. Results for 25OHD(3) are comparable to those obtained by LC-MS/MS, while results for 25OHD(2) are around half of those obtained by LC-MS/MS.
维生素 D 缺乏被认为是一个主要的健康问题，因此需要可靠的常规检测方法来测量血液样本中的维生素。在此，我们对罗氏 25-羟维生素 D 总检测法（Vitamin D Total）进行了验证。
我们分析了控制材料（2 个水平）和患者血清池（3 个水平），浓度范围为 34 至 123nmol/L，共进行了 84 次分析，在 21 天内进行了 21 天。我们还在 10 次单独的运行中分析了 5 个血清池，以验证定量下限。我们还分析了 53 对血清和 Li-肝素血浆样本。我们将 25-OH Vitamin D Total 检测法与我们的液相色谱串联质谱法（LC-MS/MS）方法进行了比较[194 个没有 25-羟维生素 D（2）（25OHD（2））的患者样本和 23 个含有 25OHD（2）的患者样本]。
在 34 和 56nmol/L 浓度下，日内 CV 分别为 4.8%和 1.9%，总 CV 分别为 8.3%和 6.1%。我们验证了制造商声明的定量下限为 22.5nmol/L。血清和血浆样本（Li-肝素）之间没有观察到显著差异。使用仅含有 25OHD（3）（无 25OHD（2））的 194 个样本进行与 LC-MS/MS 的比较显示，Vitamin D Total nmol/L=1.07×（LC-MS/MS）nmol/L+4.7 nmol/L，而使用 23 个患者样本进行 25OHD（2）的比较显示，Vitamin D Total nmol/L=0.55×（LC-MS/MS）nmol/L-2.38 nmol/L（Demings 回归）。
罗氏 Vitamin D Total 检测法被认为适合于测量血清和 Li-肝素血浆中的 25OHD。25OHD（3）的结果与 LC-MS/MS 获得的结果相当，而 25OHD（2）的结果约为 LC-MS/MS 获得的结果的一半。
