摘要
目的评估免疫组织化学和免疫荧光染色方法在肾活检组织石蜡切片磷脂酶A2受体（PLA2R）检测中的应用情况,寻找在肾组织中检测PLA2R准确、快捷的实验方法。方法对193例肾活检组织石蜡切片进行PLA2R免疫组织化学染色,抗原修复方法为高压锅热修复加胰蛋白酶双修复法,其中包括特发性膜性肾病（IMN）139例、膜型狼疮性肾炎15例、乙型肝炎病毒相关性膜性肾病8例、Ig A肾病18例和微小病变13例。将其中22例肾组织PLA2R阳性IMN患者的肾组织石蜡切片分别给予高压锅热修复、高压锅热修复加胰蛋白酶双修复、水浴热修复和水浴热修复加胰蛋白酶双修复等4种抗原修复法,然后行PLA2R免疫组织化学染色,对染色效果进行比较。再将其中15例肾组织PLA2R阳性IMN患者的肾组织石蜡切片分别给予水浴热修复加胰蛋白酶双修复、蛋白酶K修复、胃蛋白酶修复等3种抗原修复法,然后行PLA2R免疫荧光染色,对染色效果进行比较。结果 193例标本中,IMN患者PLA2R免疫组织化学染色结果阳性率为90.6%（126/139）;其他54例非IMN患者均为阴性。采用高压锅热修复加胰蛋白酶双修复法和水浴热修复加胰蛋白酶双修复时,22例患者的PLA2R免疫组织化学染色结果均为阳性;而采用高压锅热修复法或水浴热修复时,仅部分患者为阳性。采用水浴热修复加胰蛋白酶双修复、蛋白酶K修复和胃蛋白酶修复时,15例患者的PLA2R免疫荧光染色结果均为阳性,但水浴热修复加胰蛋白酶双修复为弥漫球性着色,荧光强度2＋～3＋,而蛋白酶K修复和胃蛋白酶修复为局灶节段性着色,荧光强度3＋～4＋。结论肾组织PLA2R免疫组织化学染色可有效鉴别IMN与继发性MN。行免疫组织化学染色和免疫荧光染色时,首选水浴热修复加胰蛋白酶双修复法。
Objective To evaluate the application of immunohistochemistry and fluorescence staining method in the detection of phospholipase A2 receptor（ PLA2R） on paraffin section of renal biopsy tissue,and to find an accurate and fast method for the detection of PLA2 R in renal tissue. Methods The PLA2 R of 193 caseswere detected by immunohistochemical staining,and the antigen was repaired by the method of high pressure cooker（ HPC） hot repair plus trypsin repair. The 193 samples including 139 cases of idiopathic membranous nephropathy（ IMN）,15 cases of membranous lupus nephritis,8 cases of hepatitis B virus associated membranous nephropathy,18 cases of Ig A nephropathy,and 13 cases of minimal change diseases. To compare the dyeing effects,22 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2 R were stained by using 4 different methods of antigen repairing,which included HPC hot repair,HPC hot repair plus trypsin repair,water bath heat repair,and water bath heat repair plus trypsin repair. To compare the dyeing effects,15 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2 R were stained by using 3 different methods of antigen repairing,which included water bath heat repair plus trypsin repair,protease K digestion repair,and pepsin digestion repair. Results In 193 cases,the positive rate of PLA2 R in IMN cases was 90. 6%（ 126 /139）,and the other 54 patients without IMN were negative. Twenty-two IMN patients were positive for PLA2 R by using the HPC heat repair plus trypsin repaire or the water bath heat repair plus trypsin repair; while only a few cases of 22 IMN cases were positive by using the HPC hot repair alone or water bath heat repair alone. Fifteen IMN patients were positive for PLA2 R by using water bath heat repair plus trypsin repair,protease K digestion repair,and pepsin digestion repair,but the distribution of positive deposits and the background were different. Conclusions PLA2 R immunohistochemical staining can effectively identify IMN and secondary
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2015年第5期562-566,共5页
Acta Academiae Medicinae Sinicae
基金
国家科技支撑计划课题(2011BAI10B06)~~
关键词
肾组织
磷脂酶A2受体
免疫组织化学
免疫荧光
抗原修复
renal tissues
phospholipase A2 receptor
immunohistochemistry
immunofluorescence
antigen retrieval
