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SDS-PAGE-Based Quantitative Assay for Screening of Kidney Stone Disease
oleh: Wing-Seng Leong, Wai-Hoe Lau, Ismail Zhari, Lay-Harn Gam
Format: | Article |
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Diterbitkan: | BMC 2009-01-01 |
Deskripsi
<p>Abstract</p> <p>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 (<it>P</it> < 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.</p>