Biol Proced Online. 2009 May 15;11:145-60. doi: 10.1007/s12575-009-9007-y.
Kidney stone disease is a common health problem in industrialised nations. We developed a SDS-PAGE-based method to quantify Tamm Horsfall glycoprotein (THP) for screening of kidney stone disease. Urinary proteins were extracted by using ammonium sulphate precipitation at 0.27 g salt/mL urine. The resulted pellet was dissolved in TSE buffer. Ten microliters of the urinary proteins extract was loaded and separated on 10% SDS-PAGE under reducing condition. THP migrated as single band in SDS-PAGE. The assay reproducibility and repeatability were 4.8% CV and 2.6% CV, respectively. A total of 117 healthy subjects and 58 stone patients were tested using this assay, and a distinct cut-off (P < 0.05) at 5.6 μg/mL THP concentration was used to distinguish stone patients from healthy subjects. The sensitivity and specificity of the method were 92.3% and 83.3%, respectively.
肾结石病是工业化国家常见的健康问题。我们开发了一种基于 SDS-PAGE 的方法来定量检测 Tamm-Horsfall 糖蛋白（THP），以筛查肾结石病。通过在尿液中使用 0.27 g 盐/mL 的硫酸铵沉淀来提取尿液中的蛋白质。所得的沉淀溶解在 TSE 缓冲液中。取 10 μL 尿液蛋白提取物上样，并在还原条件下于 10% SDS-PAGE 中分离。THP 在 SDS-PAGE 中迁移为单一条带。该测定的重复性和再现性分别为 4.8% CV 和 2.6% CV。使用该方法检测了 117 名健康受试者和 58 名结石患者，使用 5.6μg/mL THP 浓度的明显截止值（P<0.05）来区分结石患者和健康受试者。该方法的灵敏度和特异性分别为 92.3%和 83.3%。
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