尿液中红桂木凝集素结合蛋白的检测及临床意义

Research on artocarpus lingnanensis lectin binding proteins in human urine and its clinical significance.

目的 建立酶标记红桂木凝集素 (ALL HRP)夹心法检测尿液中ALL结合蛋白。方法 制备ALL HRP并经SephadexG 2 0 0纯化作为酶标凝集素 ,ALL为包被物 ,确定最适包被浓度及ALL HRP工作稀释度 ,夹心酶联检测尿液中ALL结合蛋白 (U LBP)。经精密度、重现性等实验并制备标准曲线。结果 以U LBP作标准制备的标准曲线线性检测范围为 9 4～ 15 0 μg/ml。本法测定几种疾病的尿U LBP/Cr( μg/ μmol) ,以SLE肾病为最高 4 4 0 6± 3 2 4 7,其次是急性肾炎 3 8 0 7± 2 6 13 ,肾病综合征 3 2 5 7± 2 0 3 3 ,血尿 2 5 3 0± 14 62 ,慢性肾炎 18 0 3± 13 70 ,糖尿病 15 69±12 0 8,与正常对照组相比 ,均有显著性差异。本法批内CV为 8 86% ,批间CV为 9 89%。结论 ALL HRP夹心酶联法检测U LBP方法简便 ,结果稳定 ,可望在临床上推广使用。

Objective\ ALL HRP Sandwich assay was developed for the measurement of Artocarpus lingnanensis lectin binding proteins in human urine(U LBP) Methods \ ALL HRP was prepared and purified by Sephadex G 200 chromatography The optimum concentration of coating ALL and the working dilution degree of ALL HRP were determined for the measurement of U LBP With Sandwich enzyme linked assay intra and inter run precision studies were performed The standard curve was prepared for U LBP Results\ The working concentrafion range of the standard curve was 9 4～150 μg/ml for U LBP The value of U LBP/Cr(mg/mmol)in normal human urine or patient′s urine with SLE nephropathy,acute or chronic glomerulonephritis(AGN or CGN),nephrotic syndrome,hematuria,pyelonephritis,diabetes mellitus,were determined with ALL HRP Sandwich enzyme linked assay respectively The value of U LBP/Cr(mg/mmol)in normal human urine was 3 44±1 56 mg/mmol,the value of U LBP/Cr(mg/mmol)in SLE nephropathy patient urine,44 06±32 47 mg/mmol was highest;secondly,AGN,38 07±26 13 mg/mmol;nephrotic syndrome 32 57±20 33 mg/mmol;hematuria 25 30±14 62 mg/mmol;pyelonephritis 17 37±11 77 mg/mmol;CGN 18 03±13 70 mg/mmol;diabetes mellitus 15 69±12 08 mg/mmol;respectively The comparison of the value of U LBP/Cr between patients and normal human was distinctly differential Intra assay CV 8 96%,inter assay CV 9 89% Conclusion\ The measurement on U LBP in urine by Sandwich enzyme linked assay with ALL HRP is simple and precise,can be used in wide range The assay on U LBP in urine is important for the diagnosis on some kidney diseases