Characterization and Clinical Significance of Natural Variability in Hepatitis B Virus Reverse Transcriptase in Treatment-Naive Chinese Patients by Sanger Sequencing and Next-Generation Sequencing
Characterization and Clinical Significance of Natural Variability in Hepatitis B Virus Reverse Transcriptase in Treatment-Naive Chinese Patients by Sanger Sequencing and Next-Generation Sequencing
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通过桑格测序和二代测序研究未接受治疗的中国患者乙型肝炎病毒逆转录酶自然变异的特征和临床意义
DOI：
10.1128/jcm.00119-19
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发表时间：
2019-08-01
影响因子：
9.4
通讯作者：
Ou, Qishui
中科院分区：
文献类型：
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作者：
Fu, Ya;Zeng, Yongbin;Ou, Qishui
乙型肝炎病毒 (HBV) 逆转录酶 (RT) 突变与长期抗病毒治疗期间的核苷类似物 (NA) 耐药性相关。然而，未经治疗的患者中 HBV RT 突变的特征尚未得到很好的说明。本研究的目的是调查初治患者 HBV RT 自然变异的特征和临床意义。摘要 乙型肝炎病毒 (HBV) 逆转录酶 (RT) 突变与长期抗病毒治疗期间的核苷类似物 (NA) 耐药相关。然而，未经治疗的患者中 HBV RT 突变的特征尚未得到很好的说明。本研究的目的是调查初治患者 HBV RT 自然变异的特征和临床意义。通过桑格测序分析了 427 名患者的 HBV RT 序列，通过下一代测序分析了 66 名患者的 HBV RT 序列。通过桑格测序，除 RT 中的 A181T (rtA181T) 外，未发现原发性或继发性 NA 抗性 (NAr) 突变，但通过二代测序检测到了这些突变。桑格测序发现 56 个 RT 氨基酸 (aa) 位点发生突变，其中 36 个突变可能导致 RT 或 S 蛋白中 B 或 T 细胞表位发生变化。 RT 区域内不同部分的突变分布不同。多种突变显示与 HBV DNA、HBsAg、HBeAg、年龄和肝纤维化严重程度显着相关。通过下一代测序，在晚期肝病 (ALD) 组中发现 rt251、rt266、rt274、rt280、rt283、rt284 和 rt286 的突变最多。本研究表明，下一代测序（NGS）比桑格测序更适合监测初治患者中低发生率的 NAr 突变，并且 RT 区域的突变可能与 ALD 的进展有关。
Mutations in hepatitis B virus (HBV) reverse transcriptase (RT) are associated with nucleos(t)ide analogue (NA) resistance during long-term antiviral treatment. However, the characterization of mutations in HBV RT in untreated patients has not yet been well illustrated. The objective of this study was to investigate the characterization and clinical significance of natural variability in HBV RT in treatment-naive patients. ABSTRACT Mutations in hepatitis B virus (HBV) reverse transcriptase (RT) are associated with nucleos(t)ide analogue (NA) resistance during long-term antiviral treatment. However, the characterization of mutations in HBV RT in untreated patients has not yet been well illustrated. The objective of this study was to investigate the characterization and clinical significance of natural variability in HBV RT in treatment-naive patients. HBV RT sequences were analyzed in 427 patients by Sanger sequencing and in 66 patients by next-generation sequencing. Primary or secondary NA resistance (NAr) mutations were not found, except A181T in RT (rtA181T) by Sanger sequencing, but they were detected by next-generation sequencing. Mutations were found in 56 RT amino acid (aa) sites by Sanger sequencing, 36 of which had mutations that could lead to changes in B or T cell epitopes in the RT or S protein. The distribution of mutations was diverse in different sections within the RT region. Multiple mutations showed significant association with HBV DNA, HBsAg, HBeAg, age, and severity of liver fibrosis. Mutations at rt251, rt266, rt274, rt280, rt283, rt284, and rt286 were found most in the advanced liver disease (ALD) group by next-generation sequencing. The present study demonstrates that next-generation sequencing (NGS) was more suitable than Sanger sequencing to monitor NAr mutations at a low rate in the treatment-naive patients, and that mutations in the RT region might be involved in the progression to ALD.