原位杂交检测石蜡包埋组织中丙型肝炎病毒RNA

DETECTION OF HEPATITIS C VIRUS RNA IN PARAFFIN-EMBEDDED LIVER TISSUES BY IN SITU HYBRIDIZATION

为了提高ＨＣＶＲＮＡ的原位杂交检出率，我们应用地高辛标记ＨＣＶ５＇非编码区ｃＤＮＡ作探针，采用原位分子杂交法对９６Ｎ肝硬变，１０２例肝细胞肝癌进行了ＨＣＶＲＮＡ检测，并结合不同年份标本中ＨＣＶＲＮＡ的检出率，探讨ＨＣＶＲＮＡ在肝脏的分布及其丢失问题．结果显示ＨＣＶＲＮＡ定位于肝细胞和肝癌细胞胞浆内，肝脏其它细胞未见明确阳性杂交信号。ＨＣＶＲＮＡ杂交阳性细胞呈灶状和弥漫分布，正常对照、替代、空白对照为阴性，在不同年份肝硬变及肝细胞肝癌中两两比较表明新近包埋蜡块组织较陈旧蜡块组织ＨＣＶ ＲＮＡ检出率明显高。本文结果证实ＨＣＶＲＮＡ存在于肝细胞和癌细胞的胞浆内，未见肝内其它细胞阳性，还发现ＨＣＶＲＮＡ在肝硬变、肝细胞癌中的检出率随保存时间的延长，其阳性检出率明显降低，表明ＨＣＶＲＮＡ在蜡块中存在明显的降解，应尽可能选用近期标本、为临床分析ＨＣＶ感染，ＨＣＶ与肝硬变、肝细胞肝癌的关系提供更客观的数据。

In order to increase the HCV RNA detection rate by in situ hybridization(ISH), we used digoxigenin labeled HCV 5' no coding region cDNA as probe to detect HCV RNA in 96 cases of liver cirrhosis and 102 cases of hepatocellular carcinoma by ISH. We also studied the distribution of HCV RNA in liver and the loss of HCV RNA in the specimens of different years.Our results showed that HCV RNA was detected in the cytoplasm of hepatocytes and cancerrous cells but not in the other cells of hepatic tissue. The HCV RNA positive cells distributed in a patchy, clustered and diffuse manner. No positive signal was found in the control.The detection rate of HCV RNA in lately embedded specimens was much higher than that of in old specimens where compared statistically by X￣x test. This suggested that there was obvious loss of HCV RNA in paraffin embedded specimens and to obtain the optimal results,fresh specimens should be used.