MacFarlane G D, Scheller D G, Ersfeld D L, Shaw L M, Venkatarmanan R, Sarkozi L, Mullins R, Fox B R
DiaSorin Inc., Stillwater, MN 55082, USA.
Clin Chem. 1999 Sep;45(9):1449-58.
The analytical validation of multiple lots of the PRO-Trac II ELISA (DiaSorin) for the determination of tacrolimus in whole blood is described.
The analytical parameters assessed included analytical sensitivity, dilution linearity, functional sensitivity, values in samples containing no tacrolimus, intra- and interassay precision, supplementation and recovery, metabolite cross-reactivity, interference studies, and method comparisons HPLC-tandem mass spectrometry (HPLC/MS/MS) and the IMx Tacrolimus II multiparticle enzyme immunoassay. Where appropriate, assessments were performed according to NCCLS guidelines.
The mean analytical detection limit was <0.25 microg/L for all lots, whereas the functional sensitivity was 1.0 microg/L. Excellent linear correlation (r = 0.985) was observed for dilution linearity. The intraassay imprecision was <7%, and the total imprecision by ANOVA was <10%. Recovery was 109% +/- 11%. Metabolite cross-reactivity was consistent with previous reports for this antibody. No interference was observed for 35 tested drugs. Method comparison with HPLC/MS/MS showed no statistically significant differences. Samples exhibited stability through four freeze/thaw cycles and for 1 week at room temperature.
These data demonstrate that the PRO-Trac II ELISA is a robust, accurate, and precise tool for the assessment and management of tacrolimus blood concentrations in transplant patients.
描述了对用于测定全血中他克莫司的多批次PRO-Trac II酶联免疫吸附测定法（DiaSorin）进行的分析验证。
评估的分析参数包括分析灵敏度、稀释线性、功能灵敏度、不含他克莫司的样品中的值、批内和批间精密度、加样回收率、代谢物交叉反应性、干扰研究以及与高效液相色谱-串联质谱法（HPLC/MS/MS）和IMx他克莫司II多颗粒酶免疫测定法的方法比较。在适当情况下，根据美国国家临床实验室标准委员会（NCCLS）指南进行评估。
所有批次的平均分析检测限均<0.25μg/L，而功能灵敏度为1.0μg/L。稀释线性观察到极佳的线性相关性（r = 0.985）。批内不精密度<7%，方差分析得出的总不精密度<10%。回收率为109%±11%。代谢物交叉反应性与该抗体先前的报告一致。35种受试药物未观察到干扰。与HPLC/MS/MS的方法比较显示无统计学显著差异。样品在四个冻融循环以及室温下保存1周时表现出稳定性。
这些数据表明，PRO-Trac II酶联免疫吸附测定法是评估和管理移植患者他克莫司血药浓度的一种可靠、准确且精密的工具。
