应用非同位素原位杂交检测肝组织中丙型肝炎病毒基因的分布  

Detection of Hepatitis C Virus RNA Intrahepatic Distribution is Patients with Hepatitis C by a Non-radioactive in Situ Hybridization

作　　者：王福生[1] 韩凤连[1] 王松山[1] 雷周云[1] 王林杰[1] 陆江春 皇甫玉珊[1] 

机构地区：[1]北京解放军302医院生物工程研究室

出　　处：《中华实验和临床病毒学杂志》1995年第4期314-317,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学(青年)基金

摘　　要：为了研究丙型肝炎患者肝组织中丙型肝炎病毒（ＨＣＶ）基因的分布，我们应用地高辛标记的ＨＣＶ基因５＇端非翻译区的探针（长度为３２个寡核苷酸），对２４例急、慢性丙型肝炎患者活检的肝组织石蜡包埋切片进行了原位核酸杂交检测。结果显示：ＨＣＶ基因阳性的肝组织标本有１１例，检出率是４５．８％（１１／２４）。ＨＣＶ基因主要分布于肝细胞浆，偶见于肝细胞核内。此外在肝血窦的ｋｕｐｆｆｅｒ细胞、小血管内皮细胞和汇管区附近均有明显的ＨＣＶ基因阳性染色，而对照组均未发现ＨＣＶ基因阳性信号。In order to investigate the hepatitis C virus(HCV) infection and its intrahepatic distribution in the liver tissue from the petients with acute or chronic hepatitis C, a synthetic complementary oligonucleotide probe(32-mer, located at the highly conserved 5' noncoding region of HCV genome) was endlabeled with digoxigenin-conjugated 11-dd-UTP. HCV RNA plus strand was defected by a non-radioactive in situ hybridization (NRSH) assay in the liver biopsy specimens of 24 inpetients(male 18, female 6, with average age 19±13.1 years old) with AH 6 cases, CAH(15 cases) and CPH (3 cases).The results showed that the HCV RNA in the liver tissue was detected in 45.8% of the total cases (11/24); On one nucleoplasm,and the staining density within the hepatocyte cytoplasm was ariable where as on the other hand,in the periportal region close to the lymphoid-like follicle and the Kupffer cells of the hepatic sinusoid,and the hepatic small vascular endothelium, the dense or diffuse or diffuse HCV RNA positive stainings were found respectively in a crisp or small piecemeal forms.Furthermore,in the controls the HCV RNA positive signals were not observed,and the HCV RNA poativc signals disappeared when the positive specimens were predigested with RNase A,and that suggests that the NRSH experimental method used in this study has a high specificity.

关 键 词：丙型肝炎病毒 基因 原位杂交 肝组织 丙型肝炎 

分 类 号：R373.2[医药卫生—病原生物学] R512.630.4[医药卫生—基础医学]