原位杂交法检测外周血单个核细胞中HCV RNA

目的比较慢性丙型肝炎患者用干扰素治疗前以及治疗后３个月ＰＢＭＣ中ＨＣＶＲＮＡ。方法应用地高辛素标记ＨＣＶＲＮＡ正链及负链探针，建立原位杂交方法检测外周血单个核细胞（ＰＭＢＣ）中的ＨＣＶＲＮＡ。结果治疗前１９例患者正链ＨＣＶＲＮＡ阳性，８例负链ＨＣＶＲＮＡ阳性，用正链探针杂交在较多的细胞中出现杂交信号，负链探针杂交仅在少数细胞中出现杂交信号，ＨＣＶＲＮＡ在ＰＢＭＣ胞浆中呈均质性分布。用干扰素治疗结束后３个月２０例患者中９例ＨＣＶＲＮＡ转阴性，近期治愈率４５％。结论原位杂交技术的敏感性及特异性较高，且重复性较好，是研究ＨＣＶＲＮＡ在组织中定位分布和病毒复制场所一种切实可行的方法

Detection of hepatitis C virus RNA in the peripheral blood mononuclear cells by in situ hybridization

Objective To detect HCV RNA in PBMC of 20 cases with chronic hepatitis C before and after interferon (IFN) treatment for 3 months.Methods Using digoxin labelled plus strand and minus strand HCV RNA probes,an in situ hybridization technique suitable for detecting HCV RNA in the peripheral blood mononuclear cells (PBMC) was established. Results Before IFN treatment,plus strand HCV RNA could be detected in 19 cases and minus strand in 8 cases,the hybridization signal was detected in more cells by plus strand probes than by minus strand probes.HCV RNA distributed in the cytoplasm of PBMC homogeneously.After 3 months of IFN treatment,HCV RNA in 9 of 20 cases became negative (45%). Conclusion The results suggested that the in situ hybridization technique had high sensitivity,specificity and repeatability,which was practical for studying the location and replication of HCV RNA.