改良免疫组化法在诊断乙型肝炎病毒相关性肾炎的应用研究  

The Applied Research of Improved Immunohistochemical Staining in the Diagnosis of Hepatitis B Virus Associated Glomerulonephritis

作　　者：彭卫华[1] 陈建[1] 黄晓清[1,2] 刘庆宏[3] 郑智勇[3] 李东良[4] 庄永泽[1] 王丽萍[1] 林沁[1] 俞国庆[1] 张勇[1] 李俊霞[1] 

机构地区：[1]南京军区福州总医院肾脏科,福州350025 [2]福建中医学院 [3]南京军区福州总医院病理科,福州350025 [4]南京军区福州总医院肝胆内科,福州350025

出　　处：《中国中西医结合肾病杂志》2009年第8期683-687,I0003,共6页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：福建省自然科学基金资助项目(No.X0750047)

摘　　要：目的:探讨改良免疫组化染色法在诊断乙型肝炎病毒相关性肾炎(HBV-GN)的应用前景。方法:改良免疫组化染色法采用高温高压+XXⅣ型蛋白酶消化的抗原修复法,行一抗和二抗两步孵育法,其中二抗与酶标多聚体相连。病人肾组织标本同时行免疫荧光染色法检测作对照。结果:(1)40例血清学HBsAg阳性患者的肾穿组织标本中,改良免疫组化染色法HBsAg阳性27例,阳性率为67.5%;HBcAg阳性7例(其中HbsAg同时阳性6例,HbsAg阴性1例),阳性率为17.5%;HbsAg、HBcAg同时阴性12例。免疫荧光染色法HBsAg阳性29例(72.5%);HBcAg阳性10例(25.0%,均同时呈HbsAg阳性);HbsAg、HBcAg同时阴性11例。(2)两种染色比较,HbsAg阳性率相近(改良组化67.5%vs荧光72.5%),其中同时阳性的有22例,同时阴性的5例,总一致率为67.5%(27/40),阳性一致率在改良免疫组化染色中为81.5%(22/27),在免疫荧光染色中为75.9%(22/29)。(3)10例血清丙氨酸转氨酶升高的患者中,改良免疫组化HbsAg染色阳性9例,免疫荧光染色阳性7例;血白蛋白<30g/L而尿Pro定量未达到>3.5g/24h的患者7例;6例病理呈膜性或/和膜增殖性肾炎(其中改良组化染色阳性5例,荧光染色阳性3例)。有2例行肝组织穿刺活检,病理显示肝脏病变。结论:改良免疫组化染色可提高HBV-GN的乙肝抗原检测率,有临床应用推广价值。Objective： To investigate the applications of improved inmaunohistochemical staining （IIHS） in the diagnosis of hepatitis B virus associated glomerulonephritis （HBV- GN）. Methods： IIHS using the antigen retrieval method of high temperature and high pressure added XXIV type protease digestion, incubated with the first antibody and anti-1st antibody in succession, and The latter connected with HRP- linked polymer Comparison. Kidney tissue samples of patients stained at the same time with immunofluorescent staining （IFS） method for the control. Results： （1）In 40 patients with serum HBsAg- positive, 27 cases of kidney tissues were HBsAg positive with IIHS, the positive rate was 67.5% ; HBcAg positive in 7 cases （six cases with HbsAg positive meanwhile,one case HbsAg negative）, the positive rate was 17.5% ; 12 cases were HbsAg and HBcAg negative at the same time. With IFS 29 cases showed HBsAg- positive （72： 5% ） ,and HBcAg positive in 10 cases （25.0% ,all showed HbsAg positive at the same time） ; HbsAg and HBcAg were negative in 11 patients. （2） HbsAg- positive rate with IIHS is close to IFS （67.5% vs 72.5 % ）, 22 cases were tested HbsAg - positive with IIHS and IFS simultaneously, five cases both HbsAg - negative; the HbsAg positive coincidence rate in IIHS is 81.5% （22/27）, while 75.9% （22/29） in IFS. （3）10 patients represented serum alanine aminotransferase elevating, among them 9 cases showed HbsAg positive with IIHS and 7 cases with IFS; pathology of 6 cases was membranous and/or mernbranoproliferative glomerulonephritis （ five cases HbsAg - positive with, IIHS and three cases with IFS） ; serum albumin〈 30 g/L and 24 h urine protein quantitation〈 3.5 g in 7 cases of patients,2 patients of them were performed routine liver tissue biopsy, pathology showed liver lesions. Conclusion：Improved immunohistochemical staining can promote the antigen positive rate of hepatitis B virus in HBV-GN, Indicating the popularized value of clinical application.

关 键 词：乙型肝炎病毒相关性肾炎 改良免疫组化染色 免疫荧光染色 

分 类 号：R512.62[医药卫生—内科学] R692.3[医药卫生—临床医学]