地高辛标记探针原位杂交检测肝组织中HCV RNA

PREPARATION OF DIGOXIGENIN LABELLED PROBE AND DETECTION OF HEPATITS C VIRUS RNA IN LIVER

用地高辛标记ＨＣＶ５＇－ＮＣ区ｃＤＮＡ探针原位杂交，检测了２４例ＨＣＶ感染的慢性肝病患者肝组织中ＨＣＶＲＮＡ。结果：１７例同时检测血清抗－ＨＣＶ和ＨＣＶＲＮＡ均阳性患者中，１４例（８２．３％）肝组织检出ＨＣＶＲＮＡ。７例仅有抗－ＨＣＶ阳性患者，肝组织中未检出ＨＣＶＲＮＡ。ＨＣＶＲＮＡ特异性信号主要位于肝细胞浆。感染ＨＣＶ的肝细胞呈散在、灶状或弥漫形式分布，与ＡＬＴ水平、病理改变没有明显联系。感染ＨＣＶ肝细胞周围见较多的淋巴细胞和单核细胞浸润。结果提示ＨＣＶ在肝细胞浆复制，诱导宿主免疫反应，在慢性ＨＣＶ感染的发病机制中。

onisotopic in situ prhybridization using a digoxigenin labelled cDNA probe to5'-NC region of hepatitis C virus was applied in the detection of HCV RNA in livertissue of 24 HCV infected patients with chronic liver disease. The results showedthat HCV RNA was detected in the liver tissue from 14(82.3%)of 17 patients whowere positive for anti-HCV antibody by ELISA and HCV RNA by nested-PCR. HCVRNA was undetedtable in 7 cases who were positive only for sera antibody to HCV.The specific signals of HCV RNA were mainly localized in the cytoplasm of hepato-cyte. The distributions of the HCV infected hepatocytes were characterized as spotty,orscattered,with no apparent relation to ALT of level or pathologic changes of liver.Significant amount of lymphocytes and monocytes were observed in the periphery ofHCV infected liver cells. The results suggested that replication of HCV in the cytop-lasm of hepatocytes induce the hosts immune response to infected cells,which playsan important role in the pathogenesis of the disease.