title: Deep sequencing of hepatitis B virus basal core promoter and precore mutants in HBeAg-positive chronic hepatitis B patients.
abstract: Mutants in the basal core promoter (BCP) and precore (PC) regions of hepatitis B virus (HBV) genome are associated with the progression of chronic hepatitis B (CHB) infection. However, quasispecies characteristics of naturally occurring mutants in those regions in HBeAg-positive CHB patients has not been well described, partly limited by quantitative assay. This study aimed to develop an Ion Torrent deep sequencing assay to determine BCP and PC mutant percentages in HBeAg-positive CHB patients who were treatment naïve and correlate them with different viral and host factors. Our results showed that Ion Torrent deep sequencing could achieve high accuracy (R(2)>0.99) within a dynamic range between 1% and 100%. Twelve hotspots with prevalence of greater than 20% were observed in EnhII/BCP/PC regions. G1719T, T1753V, A1762T and G1764A were genotype C related. BCP A1762T/G1764A double mutants were generally accompanied with PC 1896 wild type or lower PC G1896A mutant percentage. Lower serum HBeAg and HBsAg levels were associated with higher BCP A1762T/G1764A mutant percentages (≥ 50%). ALT levels were higher in patients with PC G1896A mutant percentage greater than 10%. In conclusion, deep sequencing such as Ion Torrent sequencing could accurately quantify HBV mutants for providing clinical relevant information during HBV infection.
authors.affiliation: Department of Immunology, and Jiangsu Key Laboratory of Medical Science and Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang 212013, Jiangsu, China.|Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Linlin
authors.initials: L
authors.lastname: Yan
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Henghui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Ma
authors.affiliation: Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
authors.forename: Di
authors.initials: D
authors.lastname: Liu
authors.affiliation: Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, 100871, China.
authors.forename: Yulin
authors.initials: Y
authors.lastname: Kang
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Ruifeng
authors.initials: R
authors.lastname: Yang
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Jianghua
authors.initials: J
authors.lastname: Wang
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Gaixia
authors.initials: G
authors.lastname: He
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Xingwang
authors.initials: X
authors.lastname: Xie
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Lai
authors.initials: L
authors.lastname: Wei
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, 100871, China.
authors.forename: Zuhong
authors.initials: Z
authors.lastname: Lu
authors.affiliation: Department of Immunology, and Jiangsu Key Laboratory of Medical Science and Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang 212013, Jiangsu, China.
authors.forename: Qixiang
authors.initials: Q
authors.lastname: Shao
authors.affiliation: Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, China.
authors.forename: Hongsong
authors.initials: H
authors.lastname: Chen
mesh_terms: D000328:Adult; D019943:Amino Acid Substitution; D016000:Cluster Analysis; D005260:Female; D005838:Genotype; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006513:Hepatitis B e Antigens / Q000276:immunology; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics* / Q000276:immunology; D019694:Hepatitis B, Chronic / Q000453:epidemiology / Q000276:immunology / Q000821:virology*; D059014:High-Throughput Nucleotide Sequencing; D054884:Host-Pathogen Interactions; D006801:Humans; D008297:Male; D009154:Mutation; D015995:Prevalence; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D055815:Young Adult
chemical_list: D006512:Hepatitis B Core Antigens; D006513:Hepatitis B e Antigens