Hepatitis B virus (HBV) genotypes impact treatment outcomes and disease progression. The current genotyping methods have limitations in patients with low HBV viral load. In this study, a more sensitive assay has been developed for determining the HBV genotype in HBV DNA suppressed patients. Fifty‐five serum samples from 55 chronic hepatitis B patients (HBeAg−, n = 20; HBeAg+, n = 35) across genotypes A to H with long‐term nucleos(t)ide analogs (NAs) treatment were collected. All samples had HBV DNA less than 29 IU/mL. Total nucleic acid (viral DNA and RNA) was extracted and a 341 bp amplicon located at HBV S gene overlapping with reverse transcriptase domain of polymerase (pol/RT) was amplified via real time (RT)‐nested polymerase chain reaction (PCR) followed by population sequencing. HBV genotype was determined by phylogenetic analysis. The assay successfully amplified HBV S/RT gene from 53 of 55 (96.4%) patient serum samples. Phylogenetic analysis demonstrated that the genotypes of all the 53 PCR positive samples matched the historical genotypes as determined by INNO‐LiPA or RT sequence from the corresponding baseline samples. This assay was able to accurately determine HBV genotype irrespective of baseline genotype, HBeAg status, or duration of viral suppression. The ability to determine genotype in virally suppressed patients may facilitate the evaluation of novel treatment agents for HBV in this patient population.

中文翻译：

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利用 DNA 和 RNA 测序技术，针对 HBV DNA 抑制的患者开发一种灵敏的 HBV 基因分型检测方法。


乙型肝炎病毒 (HBV) 基因型影响治疗结果和疾病进展。目前的基因分型方法对于低 HBV 病毒载量患者存在局限性。在本研究中，开发了一种更灵敏的检测方法，用于确定 HBV DNA 抑制患者的 HBV 基因型。收集了 55 名长期接受核苷（酸）类似物 (NA) 治疗的基因型 A 至 H 的慢性乙型肝炎患者（HBeAg−，n = 20；HBeAg+，n = 35）的 55 份血清样本。所有样本的 HBV DNA 均低于 29 IU/mL。提取总核酸（病毒 DNA 和 RNA），并通过实时 (RT) 嵌套聚合酶链式反应 (PCR) 扩增位于 HBV S 基因处与聚合酶逆转录酶结构域 (pol/RT) 重叠的 341 bp 扩增子，然后通过群体测序。通过系统发育分析确定 HBV 基因型。该检测成功地从 55 份患者血清样本中的 53 份 (96.4%) 中扩增了 HBV S/RT 基因。系统发育分析表明，所有 53 个 PCR 阳性样本的基因型均与相应基线样本中 INNO-LiPA 或 RT 序列确定的历史基因型相匹配。该检测能够准确确定 HBV 基因型，无论基线基因型、HBeAg 状态或病毒抑制持续时间如何。确定病毒抑制患者基因型的能力可能有助于在该患者群体中评估针对 HBV 的新型治疗药物。