免疫荧光细胞化学和免疫酶细胞化学染色方法检测IgA肾病患者尿足细胞的效果比较

Comparison of effect between immunoenzymatic celluar chemistry and immunofluorescence celluar chemistry in detecting urinary podocytes

目的：比较免疫荧光细胞化学和免疫酶细胞化学染色2种方法检测IgA肾病（IgAN）患者尿足细胞的效果，为尿足细胞检测方法的选择提供依据。方法：收集43例确诊为IgAN患者（实验组）和10例正常对照研究对象（对照组）活检前连续3d的晨尿各200mL，离心上清制成涂片。IgAN患者按Lee分级分成Lee I～LeeV组。采用抗人足细胞表面标记蛋白Podocalyxin（PCX）单克隆抗体对尿沉渣涂片分别行免疫荧光细胞化学和免疫酶细胞化学染色，荧光显微镜和普通光学显微镜下计数2组研究对象尿足细胞数；采用Spearman相关分析法分析2种方法检测IgAN患者尿足细胞数的相关性。结果：2种方法检测的对照组研究对象尿中未见足细胞，2种方法检测实验组研究对象尿中均可见足细胞；免疫荧光细胞化学法检测足细胞阳性率和检出中位数低于免疫酶细胞化学法，2种方法检测尿足细胞阳性率和中位数比较差异均有统计学意义（P〈0．05）；2种方法检查患者尿足细胞数呈明显的正相关关系（r=0．842，P〈0．01）。结论：采用免疫酶细胞化学法检测同一份尿标本的效果优于免疫荧光细胞化学法。

Objective To compare the effects of immunofluorescence and immunoenzymatic celluar chemistry in detecting the urinary podocytes of IgA nephropathy （IgAN） patients and to provide basis for selecting the detection method of urinary podocytes. Methods 200 mL morning urine samples were collected from 43 cases who diagnosed as IgAN （experiment group） and 10 cases of normal control subjects （control group） for 3 consecutives days before renal biopsy, and the smears were made from the supernatant by centrifugation. The IgAN patients were classified into Lee i --Lee V groups according to Lee degree method. Immunofluorescence and immunoenzymatic celluar chemistry with monoclonal anti-podocalyxin （PCX） antibody were performed to detect the urinary podocytes and the number of urinary podocytes was counted by fluorescence and optical microscope; the correlation between the number of urinary podoeytes detected by two kinds of methods were analyzed hy Spearman correlation analysis. Results There were no urinary podocytes of the objects in control group detected by two kinds of methods, but in experiment group they were visible; the positive rate and the median of urinary podocytes detected by immunofluorescence celluar chemistry were lower than those detected by immunoenzymatic celluar chemistry （P〈 0. 05 ）; there was a positive correlation between two kinds of methods （r=0. 842, P〈0. 01）.Conclusion Immunoenzymatic celluar chemistry chemistry in detecting the urinary podocytes.