地高辛素标记探针原位杂交法检测肝组织中庚型肝炎病毒RNA  

Detection of hepatitis G virus RNA in liver tissue using digoxigenin labelled probe by in situ hybridization

作　　者：聂青和[1] 胡大荣[2] 李梦东[2] 李玲[2] 朱永红[2] 

机构地区：[1]中国人民解放军第四军医大学唐都医院传染病科,陕西省西安市710038 [2]中国人民解放军第三军医大学西南医院全军传染病中心,重庆市400038

出　　处：《世界华人消化杂志》2000年第7期771-774,共4页World Chinese Journal of Digestology

摘　　要：目的自从建立起甲、乙、丙、丁、戊5种肝炎病毒的病原学诊断之后,仍有少部分肝炎患者的病因得不到明确,因此不少学者试图探索是否还有新型肝炎病毒的存在,并进行了大量的流行病学和实验诊断的研究,认为的确存在可经肠道外传播并引起人类肝炎的致病因子.目前关于庚型肝炎病毒(HGV/GBV-C)的致病性和组织嗜性尚无结论性资料.本文目的是研究 HGV/GBV-C RNA 在肝组织中表达并进一步探讨HGV/GBV-C 引起肝脏损害的可能机制.方法应用地高辛素标记 HGV/GBV-C cDNA 探针原位杂交法检测肝组织中庚型肝炎病毒 RNA.结果检测196例肝炎患者肝组织切片中的 HGV/GBV-CRNA 阳性率为37.24%;在血清中 HGV/GBV-C RNA 呈阳性患者肝组织中该病毒 RNA 检出率为48.94%;单一 HGV/GBV-C 感染者肝组织中检出率为58.33%;阳性信号仅为胞质型.原位杂交信号阳性细胞与肝细胞变性、瘀胆、炎性细胞浸润及细胞坏死程度等并不相关.本文原位杂交与免疫组化两种检测方法对比研究结果表明,两项技术有较高的符合率(86.74%),说明这两项技术具有相辅相成的作用.结论提示 HGV/GBV-C RNA 并不直接损害肝细胞.原位杂交和免疫组化两项技术成功地应用于石蜡包埋切片,为回顾性研究 HGV/GBV-C 的致病性及致病机制提供了有价值的实验手段.AIM To study the expression of HGV/GBV-C RNA in liver tissues and discuss the mechanisms of liver damage induced by HGV/GBV-C. METHODS The expression of HGV/GBV-C RNA in the liver tissues was detected with digoxigenin labelled probe by in situ hybridization. RESULTS HGV/GBV-C RNA was detected in 37.24% of 196 liver samples of hepatitis patients.The positive rate of HGV/GBV-C RNA in liver tissues was 48.94% in all patients with serum HGV/ GBV-C RNA and 58.33% in patients with HGV/GBV-C infection alone.Positive signal of hybridization was only seen in the cytoplasm and was not relevant to hepatocyte degeneration,cholestasis, inflammatory cell infiltration,degree of cell necrosis, etc.Comparing the results and combining the use of both nucleic acid in situ hybridization and immunohist- ochemical assays,a coincidence rate of 86.74% was obtained,suggesting that the two techniques can supplement each other in action. CONCLUSION HGV/GBV-C RNA seemed not to damage hepatocytes directly,In view of that paraffin-embedding materials could be detected successfully by both in situ hybridization and immunohistochemistry,the two methods may have a potential value in retrospective investigation of the pathogenetic mechanism of HGV/ GBV-C.

关 键 词：庚型肝炎病毒 GB病毒C型 原位杂交 

分 类 号：R450[医药卫生—治疗学]