title: Ultradeep Sequencing for Detection of Quasispecies Variants in the Major Hydrophilic Region of Hepatitis B Virus in Indonesian Patients.
abstract: Quasispecies of hepatitis B virus (HBV) with variations in the major hydrophilic region (MHR) of the HBV surface antigen (HBsAg) can evolve during infection, allowing HBV to evade neutralizing antibodies. These escape variants may contribute to chronic infections. In this study, we looked for MHR variants in HBV quasispecies using ultradeep sequencing and evaluated the relationship between these variants and clinical manifestations in infected patients. We enrolled 30 Indonesian patients with hepatitis B infection (11 with chronic hepatitis and 19 with advanced liver disease). The most common subgenotype/subtype of HBV was B3/adw (97%). The HBsAg titer was lower in patients with advanced liver disease than that in patients with chronic hepatitis. The MHR variants were grouped based on the percentage of the viral population affected: major, ≥20% of the total population; intermediate, 5% to <20%; and minor, 1% to <5%. The rates of MHR variation that were present in the major and intermediate viral population were significantly greater in patients with advanced liver disease than those in chronic patients. The most frequent MHR variants related to immune evasion in the major and intermediate populations were P120Q/T, T123A, P127T, Q129H/R, M133L/T, and G145R. The major population of MHR variants causing impaired of HBsAg secretion (e.g., G119R, Q129R, T140I, and G145R) was detected only in advanced liver disease patients. This is the first study to use ultradeep sequencing for the detection of MHR variants of HBV quasispecies in Indonesian patients. We found that a greater number of MHR variations was related to disease severity and reduced likelihood of HBsAg titer.
authors.affiliation: Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan Indonesia-Japan Collaborative Research Centre for Emerging and Reemerging Infectious Disease, Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia.
authors.forename: Laura Navika
authors.initials: LN
authors.lastname: Yamani
authors.affiliation: Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan Department of Gastroenterology, Kobe University Graduate School of Medicine, Kobe, Japan yanoyo@med.kobe-u.ac.jp.
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Yano
authors.affiliation: Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan Indonesia-Japan Collaborative Research Centre for Emerging and Reemerging Infectious Disease, Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia.
authors.forename: Takako
authors.initials: T
authors.lastname: Utsumi
authors.affiliation: Indonesia-Japan Collaborative Research Centre for Emerging and Reemerging Infectious Disease, Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia.
authors.lastname: Juniastuti
authors.affiliation: Department of Gastroenterology and Hepatology, Hajj General Hospital, Surabaya, Indonesia.
authors.forename: Hadi
authors.initials: H
authors.lastname: Wandono
authors.affiliation: Department of Internal Medicine, Dr. Mohammad Soewandhie General Hospital, Surabaya, Indonesia.
authors.forename: Doddy
authors.initials: D
authors.lastname: Widjanarko
authors.affiliation: Department of Internal Medicine, Dr. Mohammad Soewandhie General Hospital, Surabaya, Indonesia.
authors.forename: Ari
authors.initials: A
authors.lastname: Triantanoe
authors.affiliation: Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan Department of Physiology, Faculty of Medicine, Dr. Sardjito Hospital, Gadjah Mada University, Yogyakarta, Indonesia.
authors.forename: Widya
authors.initials: W
authors.lastname: Wasityastuti
authors.affiliation: Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Yujiao
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Department of Gastroenterology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Rina
authors.initials: R
authors.lastname: Okada
authors.affiliation: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Japan.
authors.forename: Toshihito
authors.initials: T
authors.lastname: Tanahashi
authors.affiliation: Department of Hepatology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Murakami
authors.affiliation: Department of Gastroenterology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Azuma
authors.affiliation: Indonesia-Japan Collaborative Research Centre for Emerging and Reemerging Infectious Disease, Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia.
authors.lastname: Soetjipto
authors.affiliation: Indonesia-Japan Collaborative Research Centre for Emerging and Reemerging Infectious Disease, Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia.
authors.forename: Maria Inge
authors.initials: MI
authors.lastname: Lusida
authors.affiliation: Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Yoshitake
authors.initials: Y
authors.lastname: Hayashi
mesh_terms: D000328:Adult; D000368:Aged; D019943:Amino Acid Substitution; D005260:Female; D005787:Gene Frequency; D014644:Genetic Variation; D006509:Hepatitis B / Q000821:virology*; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007214:Indonesia; D008297:Male; D008875:Middle Aged; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D006514:Hepatitis B Surface Antigens