This study established a new polymerase spiral reaction (PSR) that combines with reverse transcription reactions for HCV detection targeting 5'UTR gene. To avoid cross-contamination of aerosols, an isothermal amplification tube (IAT), as a separate containment control, was used to judge the result. After optimizing the RT-PSR reaction system, its effectiveness and specificity were tested against 15 different virus strains which included 8 that were HCV positive and 7 as non-HCV controls. The results showed that the RT-PSR assay effectively detected all 8 HCV strains, and no false positives were found among the 7 non-HCV strains. The detection limit of our RT-PSR assay is comparable to the real-time RT-PCR, but is more sensitive than the RT-LAMP. The established RT-PSR assay was further evaluated for detection of HCV in clinical blood samples, and the resulting 80.25% detection rate demonstrated better or similar effectiveness compared to the RT-LAMP (79.63%) and real-time RT-PCR (80.25%). Overall, the results showed that the RT-PSR assay offers high specificity and sensitivity for HCV detection with great potential for screening HCV in clinical blood samples.

中文翻译：

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使用逆转录酶聚合酶螺旋反应快速灵敏地检测临床血样中的丙型肝炎病毒。

本研究建立了一种新的聚合酶螺旋反应 (PSR)，它结合逆转录反应用于针对 5'UTR 基因的 HCV 检测。为了避免气溶胶的交叉污染，使用等温放大管（IAT）作为单独的密封控制来判断结果。在优化 RT-PSR 反应体系后，其有效性和特异性针对 15 种不同的病毒株进行了测试，其中 8 种为 HCV 阳性，7​​ 种为非 HCV 对照。结果表明，RT-PSR 检测可有效检测所有 8 株 HCV 毒株，并且在 7 株非 HCV 毒株中未发现假阳性。我们的 RT-PSR 检测的检测限与实时 RT-PCR 相当，但比 RT-LAMP 更灵敏。已建立的 RT-PSR 测定法进一步评估用于检测临床血液样本中的 HCV，与 RT-LAMP (79.63%) 和实时 RT-PCR (80.25%) 相比，由此产生的 80.25% 检出率显示出更好或相似的效果。总的来说，结果表明，RT-PSR 测定法为 HCV 检测提供了高特异性和灵敏度，具有在临床血液样本中筛查 HCV 的巨大潜力。