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

Biological Procedures Online

Gam Lay-Harn¹  Zhari Ismail¹  Leong Wing-Seng²  Lau Wai-Hoe¹ 

[1]School of Pharmaceutical Sciences, Universiti Sains Malaysia, USM, 11800, Penang, Malaysia

[2]Lam Wah Ee Hospital, Penang, Malaysia

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

Others  :  797169
DOI  :  10.1007/s12575-009-9007-y

 received in 2009-01-23, accepted in 2009-04-01,  发布年份 2009

【 摘 要 】

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】

   
2009 Wai-Hoe et al.

【 图 表 】

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