血尿对十二烷基硫酸钠-尿蛋白电泳检测结果的影响  

Effects of Hematuria on the Results of Sodium Dodecyl Sulfate- Agarose Gel Urinary Protein Electrophoresis

作　　者：安崇文[1] 王思思 逄璐 许冬[1] 李海霞[1] AN Chongwen;WANG Sisi;PANG Lu;XU Dong;LI Haixia(The Department of Clinical Laboratory,First Hospital of Peking University,Beijing 100034,China)

机构地区：[1]北京大学第一医院检验科,北京100034

出　　处：《标记免疫分析与临床》2023年第2期332-337,共6页Labeled Immunoassays and Clinical Medicine

摘　　要：目的 研究血尿样本中红细胞及血红蛋白对十二烷基硫酸钠-琼脂糖凝胶蛋白电泳(sodium dodecyl sulfate-agarose gel electrophoresis, SDS-AGE)检测结果的影响。方法 制备不同程度的血尿样本,采用SDS-AGE技术进行检测,观察不同程度血尿样本中血红蛋白在凝胶片上出现的区域,并分析对检测结果的影响。结果 对尿隐血定性试验微量、1+、2+、3+及3+以上不同程度的肉眼血红蛋白尿、红细胞尿、血红蛋白水溶液等样本进行检测,发现尿隐血试验在3+以下时,对SDS尿蛋白电泳图谱不受影响,而当尿隐血试验在3+或3+以上肉眼血尿时,均出现了血红蛋白降解产物的干扰条带,通过与分子量质控品的Marker区带比较,血红蛋白的干扰区带主要出现在靠近阳极端小分子蛋白区域,即大部分在凝胶片的溶菌酶区域,此区域占比数量最多,其次少部分在磷酸丙糖异构酶区域偏向阴极端(即游离轻链单体与α1微球蛋白之间)以及白蛋白区域,分别为血红蛋白珠蛋白单体、二聚体和四聚体。对肾穿病理诊断为肾小球疾病的蛋白尿合并肉眼血尿样本及严重血管内溶血的溢出性血红蛋白尿样本进行检测,结果显示血红蛋白珠蛋白单体、二聚体会增加小分子蛋白组分的百分比,珠蛋白四聚体会增加白蛋白组分的百分比。结论 尿隐血试验在3+或肉眼血尿的样本,尿中含有的红细胞或血红蛋白(包含血红蛋白降解产物,如血红蛋白单体-珠蛋白、二聚体等)会导致SDS尿蛋白电泳检测结果受到影响,主要是对小分子蛋白区域影响较大,中分子蛋白区域影响较小,随着血红蛋白含量的增多而受影响的程度也越大,提示我们应在报告中给予注明,或将血红蛋白降解成份单独报告,避免产生对报告的错误解读。Objective To study the effects of red blood cells and hemoglobin in hematuria samples on the results of sodium dodecyl sulfate-agarose gel electrophoresis(SDS-AGE).Methods We prepared hematuria samples of different degrees,and then used SDS-AGE technology to detect the areas where hemoglobin appeared on gel tablets in hematuria samples of different degrees,and analyzed their impacts on the detection results.Results The samples of macroscopic hemoglobinuria,erythrocyte urine and hemoglobin aqueous solution with microscale,1+,2+,3+and above 3+in the qualitative test of urinary occult blood were detected.It was found that when the urinary occult blood test was below 3+,the result of SDS urinary protein electrophoresis was not affected,while when the urinary occult blood test was 3+or macroscopic hemoglobinuria of above 3+,the interference bands of hemoglobin degradation products appeared.By comparing with the marker band of molecular weight quality control,the interference zone of hemoglobin mainly appeared in the small molecular protein region near the anode end,mainly in lysozyme area of gel tablets.This area accounted for the largest number.Secondly,a small number of them tended to be the negative extreme in the region of phosphotriose isomerase(i.e.between free light chain monomers andα1-microglobulins),and albumin regions,which were hemoglobin globin monomer,dimer and tetramer.Through the detection of proteinuria combined with gross hematuria samples diagnosed as glomerular disease by renal puncture pathology and overflow hemoglobinuria samples with severe intravascular hemolysis,the results showed that hemoglobin globin monomer and dimerization increased the percentage of small molecular protein components,and globin tetramerization increased the percentage of albumin components.Conclusion For the urine occult blood test in 3+or gross hematuria samples,the red blood cells or hemoglobin contained in urine(including hemoglobin degradation products:such as hemoglobin monomer globin and dimer)will affect the resu

关 键 词：血尿 蛋白尿 血红蛋白尿 十二烷基硫酸钠 尿蛋白电泳 

分 类 号：R446.12[医药卫生—诊断学]