应用原位逆转录PCR技术检测石蜡切片中HCVRNA

DETECTION OF HEPATITIS C VIRUS RNA IN PARAFFIN SECTIONS BY IN SITU REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION TECHNIQUE

利用原位逆转录聚合酶链式反应（ＩＳＲｔＰＣＲ）技术检测了１５例肝细胞癌及其癌旁肝组织中丙型肝炎病毒（ＨＣＶ）ＲＮＡ的定位及分布，并探讨影响实验结果的重要因素。结果表明ＨＣＶＲＮＡ阳性信号主要位于肝细胞和癌细胞胞浆，胞核几乎阴性。影响结果的主要因素包括所用蛋白酶Ｋ浓度，消化切片的时间。

The technique of in situ reverse transcription polymerase chain reaction (IS Rt PCR) was used for detecting distribution and localization of hepatitis C virus RNA in 15 hepatocellular carcinoma, the surrounding liver tissue, and the important factors which affected the results of in situ Rt PCR were investigated. The results showed that positive signal of HCV RNA was mainly found in the cytoplasm of the hepatocytes and cancer cells, while HCV RNA was nearly negative in the nuclei. The important factors which affected the results of in situ Rt-PCR included appropriate concentration and digestive time of proteinase K, the choice of fixative for fixation of tissue, and the proper cycles of PCR.