肾组织石蜡切片免疫荧光技术中抗原修复方法的探讨  

Study on the methods of antigen retrieval in paraffin section of kidney biopsy

作　　者：周雅丽[1] 李远[1] 郭佳[2] 邢国兰[1,2] 刘章锁[1,2] Zhou Yali;Li Yuan;Guo Jia;Xing Guolan;Liu Zhangsuo(Centre of Renal Pathology,the First Affiliated Hospital of Zheng Zhou University,Zhengzhou 450052,China;Department of Nephrology,the First Affiliated Hospital of Zheng Zhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院肾脏病理中心,郑州450052 [2]郑州大学第一附属医院肾内科,郑州450052

出　　处：《中华肾脏病杂志》2020年第3期177-182,共6页Chinese Journal of Nephrology

摘　　要：目的 比较多种抗原修复方法在肾组织石蜡切片免疫荧光技术中的应用,探讨最佳抗原修复法.方法 选取2018年6月至2019年6月期间郑州大学第一附属医院肾脏病理中心的45例肾活检组织标本为研究对象,其中狼疮肾炎、膜性肾病、IgA肾病各10例及淀粉样变性肾病15例.分别用5种抗原修复法处理各组肾组织石蜡切片.按照标本来源和抗原修复方法分为6组:对照组(冰冻切片标本)、高压热修复联合胰蛋白酶消化双修复(高压热联合胰酶)组、微波热修复联合胰蛋白酶消化双修复(微波热联合胰酶)组、高压热修复(高压热)组、微波热修复(微波热)组、胃蛋白酶消化(胃蛋白酶)组.分析和比较5种热修复抗原方法的石蜡切片与冰冻切片免疫荧光染色和半定量评分的差异.结果 高压热联合胰酶、微波热联合胰酶组肾组织石蜡切片的免疫荧光染色结果与对照组一致,与对照组比较,两组免疫荧光半定量评分的差异无统计学意义(均P> 0.05).高压热组、微波热组和胃蛋白酶组石蜡切片免疫荧光染色结果较对照组假阴性率高,免疫荧光半定量评分的差异有统计学意义(均P< 0.05).结论 高压热修复联合胰蛋白酶消化双修复法与微波热修复联合胰蛋白酶消化双修复法为石蜡切片的最佳抗原修复方法.Objective To compare the effect of various antigen retrieval methods in paraffin sections of renal biopsy tissue,and explore the best antigen retrieval method.Methods Forty-five renal biopsy specimens were collected from the First Affiliated Hospital of Zhengzhou University,including lupus nephritis(n=10),membranous nephropathy(n=10),IgA nephropathy(n=10)and amyloidosis glomerulopathy(n=15).Five retrieval methods(including high pressure thermal retrieval combined with trypsin retrieval,microwave thermal retrieval combined with trypsin retrieval,high pressure thermal retrieval,microwave thermal retrieval and gastroprotease retrieval)were used for immunofluorescence staining of paraffin sections.Renal tissue specimens were divided into six groups according to different antigen retrieval methods,frozen section specimens used as a control group.The immunofluorescence semi-quantitative scores of paraffin sections of the five heat-repairing antigen methods and frozen sections were compared.Results Immunofluorescence staining of hyperbaric thermal retrieval combined with trypsin retrieval group and microwave thermal retrieval combined with trypsin retrieval group were similar with those of frozen sections.Compared with the control group,there were no significant difference in the semi-quantitative immunofluorescence scores between the two groups(all P>0.05).However,Immunofluorescence staining of hyperbaric thermal retrieval,microwave thermal retrieval,pepsin digestion had significantly higher false negative rate than those of frozen sections.Compared with the control group,the difference in semi-quantitative immunofluorescence score was statistically significant(all P<0.05).Conclusion High pressure heat retrieval combined with trypsin retrieval or microwave heat retrieval combined with trypsin retrieval is the first choice of antigen retrieval methods.

关 键 词：活组织检查 石蜡包埋 荧光抗体技术 抗原修复 

分 类 号：R692[医药卫生—泌尿科学]