肾组织石蜡切片Ⅳ型胶原α链免疫组织化学染色在诊断Alport综合征中的应用

Immunohistochemical staining of type Ⅳ collagen α chain in paraffin sections of renal tissue in the diagnosis of Alport syndrome

目的对Alport综合征(AS)患者肾脏穿刺组织进行石蜡切片Ⅳ型胶原α链免疫组织化学检测,探讨其临床应用价值。方法选取2018年11月至2022年7月浙江大学医学院附属第一医院肾脏病中心确诊的10例AS患者(AS病例组)、10例心脏死亡器官捐献者(DCD)供肾及10例确诊为免疫球蛋白A肾病患者(对照组)的肾组织进行石蜡切片Ⅳ型胶原α1、α3、α5链全自动免疫组织化学染色,将其与冰冻切片免疫荧光染色结果进行比较。将对照组肾组织石蜡切片分别采用多种方法进行抗原修复,比较修复效果。结果对照组肾组织2种染色方法Ⅳ型胶原α1、α3、α5链染色在肾小球基底膜(GBM)中均为连续线状阳性表达。AS病例组肾组织2种染色方法Ⅳ型胶原α1链染色在GBM均呈连续线状阳性表达,α3、α5链染色在GBM为阴性或节段弱阳性表达。2种染色方法染色结果具有较高的一致性(Kappa=0.615,P=0.035)。结论石蜡切片全自动免疫组织化学检测Ⅳ型胶原α1、α3和α5能较好地用于AS的诊断,为AS的诊断和研究提供了一种可靠的技术手段。

Objective Immunohistochemical detection of type Ⅳ collagen alpha chain in paraffin sections of renal puncture tissues from patients with Alport syndrome(AS)was performed to explore its clinical application.Methods Kidney tissues from 10 AS patients(AS case group),10 cardiac death organ donor(DCD)donor kidneys and 10 patients diagnosed with immunoglobulin A nephropathy(control group)diagnosed in Kidney Disease Center,the First Affiliated Hospital of Zhejiang University School of Medicine from November 2018 to July 2022 were selected for fully automated immunohistochemical staining of type Ⅳ collagen α1,α3 and α5 chains in paraffin sections,which were compared with the results of immunofluorescence staining of frozen sections.The paraffin sections of renal tissues from the control group were subjected to antigenic repair by various methods respectively,and the repair effects were compared.Results In the control group,the two staining methods of renal tissue,type Ⅳ collagenα1,α3 andα5 chains staining,showed continuous linear positive expression in the glomerular basement membrane(GBM).The two staining methods of renal tissue of the AS case group.Type Ⅳ collagenα1 chain staining,showed continuous linear positive expression in the GBM,andα3,α5 chains staining showed negative or weakly positive expression in the GBM.T he staining results of the two staining methods were in high consistency(Kappa=0.615,P=0.035).Conclusion Fully automated immunohistochemical detection of type Ⅳ collagen α1,α3 and α5 in paraffin sections can be used for diagnosis of AS better,which provides a reliable technique for the diagnosis and study of AS.