特异核酸捕获RT-PCR技术建立及在HCV RNA检测中的应用

目的 解决ＲＴ－ＰＣＲ检测ＨＣＶＲＮＡ方法的重复性较差问题，使其成为常规实验室检测方法。方法 采用亲合素生物素系统将ＨＣＶ特异的核酸片段包被在微孔板上，利用特异核酸捕获样本中的ＨＣＶＲＮＡ然后进行逆转录和ＰＣＲ扩增，从而建立了特异核酸捕获ＲＴ－ＰＣＲ检测ＨＣＶＲＮＡ方法，并与酶消化法和抽提法进行比较，评价其敏感性、特异性、重复怀及抗污染能力。结果 特异性核酸获ＲＴ－ＰＣＲ获得的电要带较为清晰，其特

Detection of HCV RNA Using Specific Nuclear Acid Captured Reverse Transcription-polymerase Chain Reaction

Objective To solve the poor reproducibility of RT--PCR method in detection of HCV RNA in order to make it a routine laboratory assay. Methods Specific nuclear acid captured RT--PCR was established through capturing HCV RNA to microplate using specific nuclear acid employed avidin--biotin system, and amplifying captured HCV RNA thereafter. The sensitivity, specificity, reproducibility and the ability against contamination were evaluated, and compared with enzyme digestion method and extraction method. Results Specific nuclear acid captured RT--PCR could gain sharper electrophoresis bands. There was no significant difference in sesitivity and specificity as compared with enzyme digestion method and extraction method. But the reproducibility was better than that of extraction methods, and the ability against contamination was better than that of both enzyme digestion method and extraction method. Conclusion Specific nuclear acid captured PCR could promote the sharpness of clectrophoresis bands and efficiently discharge most of non--specific nuclear acids. And it possessed wonderful ability againot contamination. Thus, specific nuclear acid captured RT--PCR will be a very useful assay in the future.