乙型肝炎病毒(HBV)相关性肾炎患者肾活检组织中HBV DNA分布的分子生物学检测

为探讨乙型肝炎病毒（ＨＢＶ）相关性肾炎的发病机理，应用地高辛素标记ＨＢＶＤＮＡ探针原位分子杂交（ＩＳＨ）和直接原位聚合酶链反应（ＩＳ－ＰＣＲ）技术，对２０例临床诊断为ＨＢＶ相关性肾炎患者肾活检石蜡包埋组织切片检查。在ＩＳＨ中采用ＨＢＶＤＮＡ两种探针：用全长段探针，８５％（１７／２０）ＨＢＶＤＮＡ阳性，这１７例阳性肾组织切片再用ＨＢＶＤＮＡＳ加Ｃ段探针检测，１４例阳性（８２．３５％）。在ＩＳ－ＰＣＲ中本组病例８５％（１７／２０）亦阳性。发现ＨＢＶＤＮＡ阳性颗粒弥漫沉积于肾小球毛细血管袢、系膜区、肾小管，表现形式以浆核型、核型为主。同组病例肾组织免疫组化染色法（ＬＳＡＢ）显示，ＨＢｓＡｇ与ＨＢＶＤＮＡ的存在部位基本一致。提示ＨＢＶ引起的肾脏病变不仅是免疫介导的损害，亦可能有病毒侵犯参加免疫反应。

Detection of hepatitis B virus (HBV) DNA in renal biopsied tissues from patients with HBV related glomerulonephritis using molecular biology techniques

The purpose of this study was to determine the pathogenesis of hepatitis B virus (HBV) related glomerulonephritits. Measurement of HBV DNA was performed in paraffin embedded renal biopsied tissues from 20 patients with HBV related glomerulonephritis by using in situ hybridization (ISH) with digoxigenin labelled probe and in situ polymerase chain reaction (IS-PCR). HBV DNA staining was detected in 17 out of 20 patients (85%) with probe of full length HBV DNA and in 14 out of 17 patients (82.35%) with a probe of S C segments of HBV DNA. 17 out of 20 patients (85%) were also positive by IS-PCR assay. HBV DNA positive particles were diffusely distributed in glomerular capillary loops, mesangial areas, renal tube, and they were mainly in the presentation of cytoplasm nucleus and nucleus. The distribution of HBsAg detected by immunohistochemical technique (LSAB assay) coincided with that of HBV DNA. It is suggested that renal lesions due to HBV are not only the results of immunologic response, but also the outcome of direct invasion of HBV to this organ. Our study provided new evidencc on the molecular mechanism level for the research of the pathogenesis in HBV related glomerulonephritis.