探讨改进的免疫荧光方法检测尿液足细胞的实用性  

Assessment of modified immunofluorescence staining method for urinary podocytes of detection

作　　者：刘颖[1] 黄海长[1] 李惊子[1] 

机构地区：[1]北京大学第一医院肾内科暨北京大学肾脏病研究所,100034

出　　处：《北京医学》2005年第11期667-669,共3页Beijing Medical Journal

基　　金：北京大学211工程-人类功能基因与疾病基因研究学科群基金资助项目(编号2004-03-11)

摘　　要：目的介绍一种改进的尿液足细胞检测方法并探讨其临床实用性。方法采用免疫荧光法,在验证足细胞抗体特异性的基础上,从制片和荧光染色两方面对我实验室原检测尿足细胞方法加以改进,并对改进前后尿足细胞检出率和平均检出足细胞数量进行比较。结果①小鼠抗人Podocalyxin(PCX)单克隆抗体仅与肾组织的肾小球足细胞及肾脏病患者尿中足细胞特异结合;②尿沉渣涂片改为甩片,沉渣中有形成分调至适度可以减少脱片;③荧光染色同时用PI复染细胞核,可显著提高足细胞的分辨率,在狼疮肾炎Ⅳ型患者中,方法改进后平均足细胞数(16.85±12.56)个/20HP,明显高于方法改进前的(3.16±3.06)个/20HP,两组比较有显著性差异(P<0.05)。结论小鼠抗人PCX抗体可用于鉴别尿足细胞,经改进的方法能明显提高足细胞检出率。Objective To improve the method for the detection of urinary podocytes. Methods The previous methods used in our laboratory for urinary podocytes of detection were modified in the sample preparation and immunofluorescence staining. The specificity of the anti-human podocalyxin antibody was confirmed before the staining procedure. Results ① The monoclonal mouse anti-human podocalyxin antibody recognized glomerular and urinary podocytes only. ②Replacements of directly coated samples by centrifuge-coated samples and adjustment for the amount of urinary sedimentation could prevent the cells from detachment. ③ Nuclear counter-staining with PI could improve the signal of positive staining. In class Ⅳ lupus nephritis patients, the mean podocalyxin positive cell numbers were significantly higher by the modified method （16.85±12.56）个/20HP] than that by the previous method （（3.16±3.06） 个/20HP] （P〈0.05）. Conclusions The monoclonal antibody against podocalyxin is suitable for identification of urinary podocytes. Detachment of samples from the glass plate is reduced, and the rate of positive staining is increased by the current method.

关 键 词：技术改进 免疫荧光方法 尿液检测 足细胞 肾脏疾病 

分 类 号：R446.6[医药卫生—诊断学]