尿视黄醇结合蛋白酶免疫测定法的建立和初步应用

An Enzyme Immunoassay for Urinary Retinol-Binding Protein and Its Preliminary Application

我们自制兔（抗人ＲＢＰ）ＩｇＧ－ＨＲＰ酶标记物，建立了一种灵敏、简便、可靠的尿视黄醇结合蛋白（ＲＢＰ）双抗体夹心酶免疫测定法。方法灵敏度为０．４ｎｇ／ｍｌ；批内Ｃ．Ｖ．６．３％，批间Ｃ．Ｖ．１４．１％；工作范围０．４～５０ｎｇ／ｍｌ。本法的１００例正常健康人（２０～７４岁，平均４６岁）的尿ＲＢＰ值，以μｇ／ｍｍｏｌＣｒ表示，其算术均数（）±标准差（ｓ）为１１．２±６．０１；几何均数和９５％位点分别为９．５和２２。１０２例尿Ａ１ｂ正常的糖尿病患者，其尿ＲＢＰ值为４５．３±１０５．７μｇ／ｍｍｏｌＣｒ，较正常人明显增高（Ｐ＜０．００１），其中４０例（占３９．３％）高于正常上限。２５例肾功能正常的慢性肾盂肾炎中有２１例（占８４％）以及２４例其它各种肾小管间质疾病中的２２例（占９１．７％），尿ＲＢＰ值超越正常上限。肾小管损害可以是糖尿病肾病的一种早期症状。尿ＲＢＰ是近曲肾小管损害的一项敏感指标，对肾小管间质疾病的早期诊断具有重要价值。

We have developed an enzyme immunoassay for urinary retinol-binding protein (RBP).The rabbit (anti hRBP) IgG-HRP conjugate was prepared in our laboratory. This assay was simple,sensitive and reliable. The working range was 0. 4 to 50ng/ml, the.detection limit 0. 04ng of RBP and the within-and between-assay C. V. were 6. 3% and 14. 1% respectively. The mean urinary excretion of RBP in 100 healthy subjects was 11. 2μg/mmol Cr.RBP excretion values were 45. 3μg/mmol Cr in 102 diabetics without microalbuminuria,40 of whom (39. 3%) exceeded the normal value. The excretion was increased significantly in diabetics as compared to controls (P<0. 001). In 21 of 25 (84% ) chronic pyelonephritis with normal renal function and 22 of 24 (91. 7% ) other tubulointerstitial diseases,the urinary excretion of RBP exceeded the normal value. The renal tubular damage may be an early symptom of diabetic nephropathy. Excretion of RBP in the urine is a sensitive index of impaired proximal tubular function. Measurement of urinary RBP can be used for the early diagnosis of tubulointerstitial diseases.