Gentilini Fabio, Dondi Francesco, Mastrorilli Cinzia, Giunti Massimo, Calzolari Claudia, Gandini Gualtiero, Mancini Danilo, Bergamini Paolo Famigli
Department of Veterinary Clinical Sciences, Alma Mater Studiorum, University of Bologna, 40064 Ozzano Emilia, Bologna, Italy.
J Vet Diagn Invest. 2005 Mar;17(2):179-83. doi: 10.1177/104063870501700214.
The aim of this study was to validate an automated immunoturbidimetric assay used to quantify human albumin in urine and to accurately measure canine albumin concentrations in both urine and cerebrospinal fluid. The partial homology existing between human and canine albumin limited the accuracy of the human assays in measuring canine albumin without method modifications. Thus, the assay was modified by calibrating the analyzer with calibrators made in the laboratory containing known concentrations of canine albumin. To prepare the set of calibrators, the albumin concentration of pooled sera of healthy dogs was assessed in 5 replicates using the BromocresolGreen assay. Pooled samples were aliquoted and serially diluted to obtain the expected concentrations of albumin (0.5, 1, 5, 13, and 30 mg/dl) for establishing the canine calibration curve. Thereafter, the performance was assessed by analyzing canine urine and CSF The modified assay accurately quantified canine albumin in both specimens, as indicated by the following. Intra- and interassay variability was 0.92% and 2.74%, respectively; recovery was 99.66% and 99.07% in urine and 105.02% in CSF No interference was detected when hemolysate and glucose were added to urine. The test was linear within the verified range (0-225 mg/dl). These results demonstrate that the modified human albumin immunoturbidimetric assay can be a useful tool in the veterinary diagnostic laboratory. It is accurate and tends itself to automatization on chemistry analyzers.
本研究的目的是验证一种用于定量尿液中人白蛋白的自动化免疫比浊法，并准确测量尿液和脑脊液中的犬白蛋白浓度。人和犬白蛋白之间存在的部分同源性限制了未经方法修改的人用检测方法在测量犬白蛋白时的准确性。因此，通过使用实验室自制的含有已知浓度犬白蛋白的校准品对分析仪进行校准，对该检测方法进行了修改。为了制备校准品组，使用溴甲酚绿检测法对健康犬混合血清的白蛋白浓度进行了5次重复评估。将混合样品进行分装并连续稀释，以获得用于建立犬校准曲线的预期白蛋白浓度（0.5、1、5、13和30mg/dl）。此后，通过分析犬尿液和脑脊液来评估其性能。如下所示，修改后的检测方法能够准确地定量两种样本中的犬白蛋白。批内和批间变异分别为0.92%和2.74%；尿液中的回收率分别为99.66%和99.07%，脑脊液中的回收率为105.02%。向尿液中添加溶血产物和葡萄糖时未检测到干扰。该检测方法在验证范围内（0-225mg/dl）呈线性。这些结果表明，修改后的人白蛋白免疫比浊法可成为兽医诊断实验室的一种有用工具。它准确且易于在化学分析仪上实现自动化。
