尿蛋白SDS琼脂糖凝胶电泳方法建立及临床应用  

Experimental study on sodium dodecyl sulfate-agarose gel electrophoresis of urinary proteins and its clinical application

作　　者：吴惠毅[1] 孙召东[1] 刘安定[2] 仇为民[1] 赵绍林[1] 杨新玲[1] 

机构地区：[1]连云港市第一人民医院检验科,江苏连云港222002 [2]蚌埠医学院检验系

出　　处：《临床检验杂志》2004年第5期341-343,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的　建立应用于临床的尿蛋白十二烷基硫酸钠 (SDS) 琼脂糖凝胶 (AGE)电泳方法。方法　分别对Ⅰ、Ⅱ、Ⅲ、Ⅳ型 4种琼脂糖、不同的缓冲对、电压和电泳时间进行实验 ,确定最佳的分离条件 ,建立测定方法。结果　当Ⅳ型琼脂糖凝胶浓度为4 0g/L ,SDS含量为 1g/L ,在pH 7.0的AMP 咪唑 HCl缓冲体系下进行电泳 ,可以有效地将尿蛋白按分子量大小进行分离。 5 7份临床尿蛋白阳性标本电泳结果表明 ,生理性蛋白尿 2 4例 ,中、高分子蛋白尿 5例 ,小分子蛋白尿 11例和混合型蛋白尿 17例。结论　本法具有分离效果好、操作简便、灵敏度高、成本低廉等优点 ,可以将尿蛋白按分子量大小分为小分子、中分子以及混合型 ,对肾脏疾病的受损部位和损伤程度的判断具有较高的价值 ,适于基层实验室的常规开展。Objectives To establish a method of sodium dodecyl sulfate-agarosegel electrophoresis(SDS-AGE) to detect the urine protein.Method We tested different experimental conditions,e.g.,types of agarose,kinds of buffer,electric voltage and length of electrophoresis time-course to obtain the optimum separation.By the established method 57 positive urine protein samples were determined.Result When the type Ⅳ agarose gel concentration was 40g/L,the SDS content was 0.1% and the AMP-Imidazole-HCl electrode buffer was pH 7.0,the urine proteins were separated effectively according to molecular weight.In terms of the results 57 urine samples were divided into 4 groups:moderate molecular protein (n=24),moderate and high molecular protein (n=5),low molecular protein (n=11),and mixed molecular protein (n=17).Conclusion The established method is low-cost,simple and highly sensitive.The urine protein can be divided by molecular weight into 4 groups:1.moderate molecular protein,2.moderate and high molecular protein,3.low molecular protein and 4.mixed protein,so the method is potentially valuable in judging the damaged part and degree of the kidney and is suitable for the grass-roots laboratory to carry out as a routine.

关 键 词：尿蛋白 临床应用 蛋白尿 SDS 琼脂糖凝胶电泳 阳性标本 中分子 小分子 AMP 大小 

分 类 号：R446[医药卫生—诊断学] R692[医药卫生—临床医学]