SDS-PAGE-Based Quantitative Assay for Screening of Kidney Stone Disease

Biological Procedures Online

关键词: Kidney stone disease;    SDS-PAGE;    Tamm Horsfall Protein and diagnostic device;   

DOI  :  10.1007/s12575-009-9007-y

来源: DOAJ

【 摘 要 】

Abstract

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.

【 授权许可】

Unknown