Liao Hao, Zhang He, Shao Jinman, Li Xiaoyong, Zheng Wei V, Li Le, Yu Guangxin, Si Lanlan, Zhou Tao, Yao Zengtao, Dai Jiuzeng, Xu Dongping, Cheng Guanxun, Qu Jiuxin, Liu Yan, Chen Junhui, Lu Fengmin
Intervention and Cell Therapy Center, Peking University Shenzhen Hospital, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, China.
Department of Clinical Laboratory, Shenzhen Third People's Hospital, National Clinical Research Center for Infectious, Southern University of Science and Technology, Shenzhen, China.
J Med Virol. 2023 Mar;95(3):e28612. doi: 10.1002/jmv.28612.
Serum hepatitis B virus (HBV) RNA is a new serological indicator reflecting viral replication with good clinical application prospects. This study aimed to clarify the dynamic changes of serum HBV RNA levels and the quasispecies of HBV RNA virus-like particles in nucleos(t)ide analogues (NAs)-experienced chronic hepatitis B (CHB) patients harboring NAs-resistant mutations and their identifiable effects on NAs resistance. We included CHB patients who were on long-term NAs treatment and with HBV DNA rebound. The longitudinally dynamics of serum HBV RNA levels were quantitatively detected, and the quasispecies differences between serum HBV DNA and serum HBV RNA were compared by high-throughput sequencing. The effect of NAs concentration pressure on altering the resistance mutations quasispecies proportion of HBV DNA and HBV RNA in cell supernatant was analyzed in vitro. A total of 447 serum samples from 36 CHB patients treated with NAs were collected. The median follow-up period was 47 months (about 4 years), and the longest follow-up period was 117 months (about 10 years). Our results showed that HBV RNA could reflect virological breakthrough in 23 (64%, 23/36) patients, and serum HBV RNA rebound earlier than HBV DNA in 12 (52%, 12/23) patients. However, serum HBV RNA remained at a consistently high level and did not fluctuate significantly with the HBV DNA rebound in 6 of 36 patients. In addition, serum HBV RNA was not consistently detectable in 7 of the 36 patients, and their serum HBV RNA was undetectable even after HBV DNA had rebounded. The proportion of drug-resistant mutations in HBV DNA was higher than that of HBV RNA by high-throughput sequencing. The results of in vitro experiments showed that the viral strains with drug-resistant mutation in HBV DNA in cell supernatants gradually become the dominant strains with the increase of NAs concentrations. Serum HBV RNA levels can reflect virological breakthrough in most NAs- treated CHB patients, but there are certain limitations. NAs alter the quasispecies composition of serum HBV DNA and serum HBV RNA, resulting in a higher detection rate of drug-resistant mutations in serum HBV DNA than in serum HBV RNA.
血清乙型肝炎病毒（HBV）RNA是一种反映病毒复制的新型血清学指标，具有良好的临床应用前景。本研究旨在阐明核苷（酸）类似物（NAs）治疗的慢性乙型肝炎（CHB）患者中，携带NAs耐药突变时血清HBV RNA水平的动态变化以及HBV RNA病毒样颗粒的准种情况，及其对NAs耐药的可识别影响。我们纳入了长期接受NAs治疗且出现HBV DNA反弹的CHB患者。定量检测血清HBV RNA水平的纵向动态变化，并通过高通量测序比较血清HBV DNA和血清HBV RNA之间的准种差异。在体外分析NAs浓度压力对改变细胞上清液中HBV DNA和HBV RNA耐药突变准种比例的影响。共收集了36例接受NAs治疗的CHB患者的447份血清样本。中位随访期为47个月（约4年），最长随访期为117个月（约10年）。我们的结果显示，HBV RNA可在23例（64%，23/36）患者中反映病毒学突破，且在12例（52%，12/23）患者中血清HBV RNA比HBV DNA更早出现反弹。然而，36例患者中有6例血清HBV RNA持续维持在高水平，且未随HBV DNA反弹而显著波动。此外，36例患者中有7例血清HBV RNA并非始终可检测到，甚至在HBV DNA反弹后其血清HBV RNA仍不可检测。通过高通量测序发现，HBV DNA中耐药突变的比例高于HBV RNA。体外实验结果显示，随着NAs浓度增加，细胞上清液中HBV DNA携带耐药突变的病毒株逐渐成为优势株。血清HBV RNA水平可在大多数接受NAs治疗的CHB患者中反映病毒学突破，但存在一定局限性。NAs改变血清HBV DNA和血清HBV RNA的准种组成，导致血清HBV DNA中耐药突变的检出率高于血清HBV RNA。
