Virology Unit, National Institute for Infectious Diseases, INMI Lazzaro Spallanzani IRCCS, Via Portuense 292, 00149 Rome, Italy.
Infectious Disease-Clinical Department, National Institute for Infectious Diseases, INMI Lazzaro Spallanzani IRCCS, Via Portuense 292, 00149 Rome, Italy.
Viruses. 2021 Jun 16;13(6):1151. doi: 10.3390/v13061151.
The risk of hepatitis C virus (HCV) recurrence after direct-acting antiviral (DAA) treatment is <0.5%. However, the distinction between HCV RNA late relapse and reinfection still represents a challenge in virological diagnostics. The aim of this study was to employ next-generation sequencing (NGS) to investigate HCV RNA recurrence in patients achieving a sustained virologic response (SVR) at least six months post-treatment. NGS was performed on plasma samples from six HCV-positive patients (Pt1-6) treated with DAA. NGS of HCV NS5B was analyzed before treatment (T0), after HCV RNA rebound (T1), and, for Pt3, after a second rebound (T2). Reinfection was confirmed for Pt5, and for the first rebound observed in Pt3. Conversely, viral relapse was observed when comparing T0 and T1 for Pt6 and T1 and T2 for Pt3. Z-scores were calculated and used to predict whether HCV-positive patient samples at different time points belonged to the same quasispecies population. A low -score of <2.58 confirmed that viral quasispecies detected at T0 and T1 were closely related for both Pt1 and Pt2, while the Z-score for Pt4 was suggestive of possible reinfection. NGS data analyses indicate that the Z-score may be a useful parameter for distinguishing late relapse from reinfection.
直接作用抗病毒药物（DAA）治疗后丙型肝炎病毒（HCV）复发的风险<0.5%。然而，HCV RNA 晚期复发与再感染之间的区别仍然是病毒学诊断中的一个挑战。本研究旨在使用下一代测序（NGS）技术来研究至少在治疗后六个月达到持续病毒学应答（SVR）的患者的 HCV RNA 复发情况。对六名接受 DAA 治疗的 HCV 阳性患者（Pt1-6）的血浆样本进行了 NGS。在治疗前（T0）、HCV RNA 反弹后（T1）以及 Pt3 第二次反弹后（T2）对 HCV NS5B 的 NGS 进行了分析。Pt5 被证实为再感染，Pt3 的第一次反弹也是如此。相反，当比较 Pt6 的 T0 和 T1 以及 Pt3 的 T1 和 T2 时，观察到病毒复发。计算 Z 分数并用于预测 HCV 阳性患者在不同时间点的样本是否属于同一准种群体。Z 分数<2.58 可确认 Pt1 和 Pt2 的 T0 和 T1 检测到的病毒准种密切相关，而 Pt4 的 Z 分数提示可能发生再感染。NGS 数据分析表明，Z 分数可能是区分晚期复发与再感染的有用参数。
