肾组织磷脂酶A2受体抗原免疫组织化学染色方法研究  

A study on immunohistochemical staining method of phospholipase A2 receptor antigen in renal tissue

作　　者：张能擘 赵杰 王美芳 王慧萍 ZHANG Nengbo;ZHAO Jie;WANG Meifang;WANG Huiping(Kidney Disease Center,The First Affiliated Hospital,Zhejiang University School of Medicine/Key Laboratory of Kidney Disease Prevention and Control Technology,Zhejiang Province/National Key Clinical Department of Kidney Diseases/Institute of Nephrology,Zhejiang University/Zhejiang Clinical Research Center of Kidney and Urinary System Disease,Hangzhou,Zhejiang 310000,China)

机构地区：[1]浙江大学医学院附属第一医院肾脏病中心/浙江省肾脏病防治技术研究重点实验室/国家临床重点专科/浙江大学肾脏病研究所/浙江省肾脏与泌尿系统疾病临床医学研究中心,浙江杭州310000

出　　处：《医药前沿》2024年第25期15-17,共3页Journal of Frontiers of Medicine

摘　　要：目的:探讨肾组织石蜡切片磷脂酶A2受体(PLA2R)抗原免疫组织化学染色方法。方法:选取2023年1—8月在浙江大学医学院附属第一医院肾脏病中心穿刺的269例肾活检组织石蜡切片进行PLA2R抗原免疫组织化学染色,包括特发性膜性肾病(IMN)183例、狼疮性肾炎46例、免疫球蛋白(Ig)A肾病和微小病变型肾病各20例,计算其阳性率。选取25例IMN(冷冻切片免疫荧光IgG4分型阳性且血清PLA2R抗体阳性),狼疮性肾炎、IgA肾病、微小病变型肾病各8例作为比较不同修复条件的预试验组和对照组,采用3种方法进行抗原修复,分别是乙二胺四乙酸(EDTA)碱性修复液热修复+蛋白酶K修复、EDTA碱性修复液热修复+胃蛋白酶修复、EDTA碱性修复液热修复+胰蛋白酶修复,修复后进行全自动免疫组织化学染色,比较染色效果。结果:预试验结果显示,采用EDTA碱性修复液热修复10 min+蛋白酶K 5 min做抗原修复的标本染色结果最佳,阳性染色沿肾小球毛细血管壁呈颗粒状沉积且背景清晰,对照组均为阴性。PLA2R抗原免疫组织化学染色结果显示IMN 183例阳性率为86.3%(158/183);狼疮性肾炎46例、IgA肾病20例、微小病变型肾病20例均为阴性。结论:肾组织PLA2R抗原免疫组织化学染色能为IMN的诊断提供有力依据。免疫组织化学染色时,首选EDTA碱性修复液热修复+蛋白酶K双修复。Objective To explore the immunohistochemical staining method of phospholipase A2 receptor(PLA2R)antigen in renal tissue paraffin sections.Methods Retrospective selection of 269 renal biopsy tissue paraffin sections from January to August 2023 at Nephrology Centre of the First Affiliated Hospital of Zhejiang University School of Medicine were performed for PLA2R antigen immunohistochemical staining,including 183 cases of idiopathic membranous nephropathy(IMN),46 cases of lupus nephritis,20 cases of immunoglobulin(Ig)A nephropathy,and 20 cases of minimal change disease.The positivity rate was calculated.A total of 25 patients with IMN(immunofluorescence IgG4 positive and serum PLA2R antibody positive),lupus nephritis,IgA nephropathy,and minimal change disease were selected as experimental and control groups to compare different antigen retrieval conditions.Three methods were used for antigen retrieval:ethylene diamine tetraacetic acid(EDTA)alkaline retrieval solution with thermal retrieval+protease K retrieval,EDTA alkaline retrieval solution with thermal retrieval+pepsin retrieval,and EDTA alkaline retrieval solution with thermal retrieval+trypsin retrieval.After retrieval,fully automated immunohistochemical staining was performed to compare the staining effects.Results Preliminary experimental results showed that specimens treated with EDTA alkaline retrieval solution for 10 minutes+protease K for 5 minutes had the best staining results for antigen retrieval.Positive staining showed granular deposition along the glomerular capillary walls with a clear background,while all samples in the control group were negative.Immunohistochemical staining of PLA2R antigen showed that the positivity rate for IMN was 86.3%(158/183).All cases of lupus nephritis,IgA nephropathy,and minimal change disease were negative.Conclusions Immunohistochemical staining of PLA2R antigen in renal tissue can provide an effective basis for the diagnosis of IMN.When performing immunohistochemical staining,the preferred method is heat retrieval w

关 键 词：特发性膜性肾病 磷脂酶A2受体 免疫组织化学染色 

分 类 号：R692[医药卫生—泌尿科学]