52 例病毒性肝炎患者肝组织中丙型肝炎病毒 RNA 和 HCAg-NS3 的测定  

Detection of HCV RNA and antigen NS3 to HCV in liver tissue by in situ hybridization and direct enzyme immuno assay

作　　者：黄德庄 闫惠平[1,2] 郎振为 张建成[1,2] 贺丽香 

机构地区：[1]北京市肝炎研究所 [2]北京佑安医院

出　　处：《中华微生物学和免疫学杂志》1997年第3期222-226,共5页Chinese Journal of Microbiology and Immunology

摘　　要：应用地高辛标记探针原位杂交法和单克隆抗ＨＣＶ－ＮＳ３－ＨＲＰ建立直接酶标免疫组化法分别测定５２例肝炎患者肝组织ＨＣＶＲＮＡ和ＨＣＡｇ－ＮＳ３。结果抗ＨＣＶ阳性组ＨＣＶＲＮＡ检出率５７．１％（１６／２８），ＨＣＡｇ－ＮＳ３检出率５３．６％（１５／２８）；抗ＨＣＶ阴性组其两项检出率均为１２．５％（３／２４）。肝组织中ＨＣＶＲＮＡ阳性物呈蓝紫色细小颗粒存在于肝细胞核或胞浆内，其在肝小叶中的分布可分为３型，即弥漫型、局灶型、散在型。肝组织中ＨＣＡｇ－ＮＳ３阳性物呈棕黄色细小颗粒分布于肝细胞核或胞浆内，以单个或数个阳性细胞散布于肝小叶中。２３例ＨＣＶＲＮＡ或／和ＨＣＡｇ－ＮＳ３阳性病例以肝炎后肝硬化（ＬＣ）病例占多数（１４／２３），其次为慢性重型肝炎（ＣＳＨ）和中度慢性肝炎（ＣＡＨ）。此两种检测方法具有较高符合率（９０．４％，４７／５２），表明病毒核酸及其表达产物均存在于肝细胞内，与ＨＣＶ感染密切相关。这为ＨＣＶ感染诊断提供了直接依据，有利于研究ＨＣＶ感染中病毒复制、慢性化进程、抗病毒治疗监测及重叠感染时病毒相互关系。We developed a nonisotopic in situ hybridization(NISH) assay to detect hepatitis C virus(HCV) RNA and a direct enzyme immuno assay(D EIA) to detect HCAg NS3 in human liver tissues.The Dig HCV cDNA probe used for NISH was labelled with PCR method to the 5′ non coding region of HCV genome.The monoclonal anti HCV NS3 was labelled with horse radish peroxidase(Sigma),52 formalin fixed paraffin embedded liver specimens from Chinese patients with acute or chronic viral hepatitis were studied.Of all 52 viral hepatitis patients,the detective rate of HCV RNA with NISH was 57 1%(16/28) in anti HCV positive group,and in anti HCV negative group 12 5%(3/24) ( P <0 01).The hybridization signals were found more in the nucleus than in the cytoplasm of the hepatocytes.The distribution of HCV RNA positive cells in hepatic lobuli were found in three patterns:patcky in 7,cluster in 9 and diffuse in 3 cases.The detective rate of HCAg NS3 was 53 6%(15/28) in anti HCV positive patients and 12 5%(3/24) in anti HCV negative patients.The positive signal was showed in small brown yellow granules in the nucleus or cytoplasm and the HCAg NS3 positive cells were distributecd sporadically in hepatic lobuli.Confirmation tests of these two methods showed well specificity.The results of these two methods were comparable,the coincident rate was 90 4%(47/52).The clinical features of 23 cases with HCV RNA and/or HCAg NS3 positive signal were discussed. Our study showed that these two techniques were highly sensitive and specific,and had more advantages on giving a direct view of viral infection with clear background for a stable longer time.Those were useful biomolecular pathological techniques for clinical and experimental studies on hepatitis C infection.

关 键 词：丙型肝炎病毒 RNA 分析 肝炎 病毒性 

分 类 号：R512.630.3[医药卫生—内科学]