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目的解决ＲＴ－ＰＣＲ检测ＨＣＶ ＲＮＡ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ＨＣＶ ＲＮＡ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ＨＣＶ ＲＮＡ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、重复性及抗污染能力。结果特异核酸捕获ＲＴ－ＰＣＲ获得的电泳条带较为清晰，其特异性、敏感性与酶消化法、抽提法无显著差异，但重复性高于抽提法，抗污染能力显著高于酶消化法和抽提法。结论特异核酸捕获ＲＴ－ＰＣＲ检测ＨＣＶ ＲＮＡ方法能有效去除非特异核酸和提高电泳条带清晰度，并具有极强的抗产物污染能力。因此，特异核酸捕获ＲＴ－ＰＣＲ具有广泛推广应用的明景。
Objective To solve the problem of poor reproducibility of RT-PCR in detecting HCV RNA, making it a routine laboratory test. Methods The avidin-biotin system was used to coat the HCV-specific nucleic acid fragments on a microplate. The specific nucleic acids were used to capture the HCV RNA in the samples and then reverse transcribed and amplified by PCR. Thus, a specific nucleic acid capture RT-PCR assay HCV RNA method, and compared with enzyme digestion and extraction method to evaluate its sensitivity, specificity, repeatability and anti-pollution ability. Results The electrophoresis band obtained by specific nucleic acid trapping RT-PCR was clear, and its specificity and sensitivity were not significantly different from enzyme digestion and extraction methods, but the repeatability was higher than that of extraction method, and the anti-contamination ability was significantly higher than that of enzyme digestion Law and extraction method. Conclusion Specific nucleic acid capture RT-PCR detection of HCV RNA method can effectively remove non-specific nucleic acid and improve the electrophoresis band clarity, and has strong resistance to product contamination. Therefore, specific nucleic acid capture RT-PCR has broad application prospects.