Department Experimental Medicine and Surgery, University of Rome "Tor Vergata", 00133, Rome, Italy.
Hepatology Unit, University Hospital of Rome "Tor Vergata", 00133, Rome, Italy.
Sci Rep. 2018 Jun 12;8(1):8988. doi: 10.1038/s41598-018-26862-y.
Natural resistance-associated substitutions (RASs) are reported with highly variable prevalence across different HCV genotypes (GTs). Frequency of natural RASs in a large Italian real-life cohort of patients infected with the 4 main HCV-GTs was investigated. NS3, NS5A and NS5B sequences were analysed in 1445 HCV-infected DAA-naïve patients. Sanger-sequencing was performed by home-made protocols on 464 GT1a, 585 GT1b, 92 GT2c, 199 GT3a, 16 GT4a and 99 GT4d samples. Overall, 20.7% (301/1455) of patients showed natural RASs, and the prevalence of multiclass-resistance was 7.3% (29/372 patients analysed). NS3-RASs were particularly common in GT1a and GT1b (45.2-10.8%, respectively), mainly due to 80K presence in GT1a (17%). Almost all GTs showed high prevalence of NS5A-RASs (range: 10.2-45.4%), and especially of 93H (5.1%). NS5A-RASs with fold-change >100x were detected in 6.8% GT1a (30H/R-31M-93C/H), 10.3% GT1b (31V-93H), 28.4% GT2c (28C-31M-93H), 8.5% GT3a (30K-93H), 45.5% GT4a (28M-30R-93H) and 3.8% GT4d (28V-30S-93H). Sofosbuvir RAS 282T was never detected, while the 159F and 316N RASs were found in GT1b (13.4-19.1%, respectively). Natural RASs are common in Italian patients infected with HCV-GTs 1-4. High prevalence of clinically-relevant RASs (such as Y93H) supports the appropriateness of HCV resistance-test to properly guide DAA-based therapy.
自然耐药相关取代（RAS）在不同 HCV 基因型（GT）中报道的流行率差异很大。本研究旨在调查意大利一个大型 HCV 感染患者的自然 RAS 频率。对 1445 例 DAA 初治 HCV 感染患者的 NS3、NS5A 和 NS5B 序列进行了分析。464 例 GT1a、585 例 GT1b、92 例 GT2c、199 例 GT3a、16 例 GT4a 和 99 例 GT4d 样本采用自制方案进行 Sanger 测序。总的来说，1455 例患者中有 20.7%（301/1455）出现自然 RAS，372 例分析患者中有 7.3%（29/372）出现多药耐药。GT1a 和 GT1b 中的 NS3-RAS 特别常见（分别为 45.2-10.8%），主要归因于 GT1a 中 80K 的存在（17%）。几乎所有 GT 都显示出 NS5A-RAS 的高流行率（范围：10.2-45.4%），特别是 93H（5.1%）。在 6.8%的 GT1a（30H/R-31M-93C/H）、10.3%的 GT1b（31V-93H）、28.4%的 GT2c（28C-31M-93H）、8.5%的 GT3a（30K-93H）、45.5%的 GT4a（28M-30R-93H）和 3.8%的 GT4d（28V-30S-93H）中检测到 NS5A-RAS 折叠变化>100x。从未检测到索非布韦 RAS282T，而 GT1b 中发现了 159F 和 316N RAS（分别为 13.4-19.1%）。意大利 HCV GT1-4 感染患者中自然 RAS 很常见。临床相关 RAS（如 Y93H）的高流行率支持 HCV 耐药性检测的合理性，以适当指导基于 DAA 的治疗。
