标本处理方法对尿液蛋白电泳的影响(1)
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[摘要] 目的 探讨不同蛋白沉淀方法对尿液标本蛋白电泳效果的影响。方法 采集尿液标本8份，分别采用丙酮沉淀法、TCA/丙酮沉淀法及ACN/TFA沉淀法对尿液标本进行蛋白沉淀，然后进行双向电泳，比较3种标本处理方法后蛋白电泳蛋白点数及图像清晰程度。结果TCA/丙酮沉淀后电泳图谱清晰，丙酮沉淀法及ACN/TFA沉淀后单边电泳图谱具有较多横纹干扰条带，尿液标本TCA/丙酮沉淀后电泳蛋白点数高于丙酮沉淀法及ACN/TFA沉淀法，差异具有统计学意义。结论 在丙酮沉淀前应用TCA可以消除干扰等电聚焦的盐类物质，进一步降低标本的导电能力，并且能够最大程度地提取尿液中的蛋白质。
[关键词] TCA/丙酮沉淀；丙酮沉淀；ACN/TFA沉淀；尿液标本；蛋白
[中图分类号] R446.1 [文献标识码] B [文章编号] 1673-9701(2011)24-131-02
Effect of Different Handling to Protein Electrophoresis on Urine Specimen
WANG Yunjie
Department of Clinical Laboratory，Aiyimei Hospital of Fenghua City in Zhejiang Provience，Fenghua 315000，China
[Abstract] Objective To investigate the effect of different handling to protein electrophoresis on urine specimen. Methods Eight urine specimen were selected and urine protein were sedimented with acetone precipitation method，Trichloroacetic acid(TCA)/ acetone precipitation method and Acetonitrile(ACN)/Trifluoroacetic acid(TFA)precipitation method，and urine specimen had a biphase electrophoresis，proteinic counting and proteinic of electrophoretogram were contrasted. Results Electrophoretogram was limpid in urine protein sedimented with TCA/acetone precipitation method，there was some transverse striation and interference strap in acetone precipitation method and ACN/TFA precipitation method，proteinic counting of electrophoretogram was higher in TCA/ acetone precipitation method than in acetone precipitation method and ACN/TFA precipitation. Conclusion TCA handling before acetone precipitation chould eliminated effected of electrofocusing salts on urine specimen，and extracted urine protein in max degree.
[Key words] TCA/ acetone precipitation；Acetone precipitation；ACN/TFA precipitation；Urine specimen；Protein
尿液是临床常见的检验标本，尿液标本容易获取，其中蛋白质含量检测是最常用的检测项目，尿液中蛋白质的含量检测能够反应肾脏及泌尿系的病变。如肾小球基底膜的滤过功能、某些泌尿系的肿瘤，甚至能够为某些泌尿系以外疾病提供诊断依据，尿液标本中含有较多蛋白外杂质成分，在检测过程中能够影响蛋白的检测效果，为此我们采用不同的方法对尿液标本中蛋白进行沉淀处理，分析其对尿液标本蛋白电泳检测的影响。
1 材料与方法
1.1 标本来源
尿液标本来自2009年12月～2010年1月本院体检中心患者常规送检尿液，均为中段尿液。所有尿液标本均在采集2h内送检。尿液标本送检后低温离心后上清液深低温冰箱保存待检。
1.2 主要仪器及试剂 ......
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