title: Different Variants in Reverse Transcriptase Domain Determined by Ultra-deep Sequencing in Treatment-naïve and Treated Indonesian Patients Infected with Hepatitis B Virus.
abstract: A nucleos(t)ide analog (NA) is the common antiviral drug available for directly treating hepatitis B virus (HBV) infection. However, its application has led to the emergence of NA-resistant mutations mostly in a conserved region of the reverse transcriptase domain of HBV polymerase. Harboring NA-resistant mutations decreases drug effectiveness and increases the frequency of end-stage liver disease. The invention of next-generation sequencing that can generate thousands of sequences from viral complex mixtures provides opportunities to detect minor changes and early viral evolution under drug stress. The present study used ultra-deep sequencing to evaluate discrepant quasispecies in the reverse transcriptase domain of HBV including NA-resistant hotspots between seven treatment-naïve Indonesian patients infected with HBV and five at the early phase of treatment. The most common sub-genotype was HBV B3 (83.34%). The substitution rate of variants determined among amino acids with a ratio of ≥ 1% changes was higher among the population in conserved regions (23.19% vs. 4.59%, P = 0.001) and in the inter-reverse transcriptase domain (23.95% vs. 2.94%, P = 0.002) in treatment naïve, than in treated patients. Nine hotspots of antiviral resistance were identified in both groups, and the mean frequency of changes in all patients was < 1%. The known rtM204I mutation was the most frequent in both groups. The lower rate of variants in HBV quasispecies in patients undergoing treatment could be associated with virus elimination and the extinction of sensitive species by NA therapy. The present findings imply that HBV quasispecies dynamically change during treatment.
authors: Division of Infectious Disease Pathology, Department of Microbiology and Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan.|Department of Physiology, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia.
authors: Widya
authors: W
authors: Wasityastuti
authors: Department of Gastroenterology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors: Yoshihiko
authors: Y
authors: Yano
authors: Department of Anatomical Pathology, Dr. Sardjito Hospital, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia.
authors: Dewiyani Indah
authors: DI
authors: Widasari
authors: Division of Infectious Disease Pathology, Department of Microbiology and Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan.
authors: Laura Navika
authors: LN
authors: Yamani
authors: Subdivision of Gastroenterohepatology, Department of Internal Medicine, Dr. Sardjito Hospital, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia.
authors: Neneng
authors: N
authors: Ratnasari
authors: Department of Anatomical Pathology, Dr. Sardjito Hospital, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia.
authors: Didik Setyo
authors: DS
authors: Heriyanto
authors: Department of Gastroenterology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors: Rina
authors: R
authors: Okada
authors: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Japan.
authors: Toshihito
authors: T
authors: Tanahashi
authors: Department of Hepatology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors: Yoshiki
authors: Y
authors: Murakami
authors: Department of Gastroenterology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors: Takeshi
authors: T
authors: Azuma
authors: Division of Infectious Disease Pathology, Department of Microbiology and Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan.
authors: Yoshitake
authors: Y
authors: Hayashi
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000998:Antiviral Agents / Q000494:pharmacology; D024882:Drug Resistance, Viral / Q000235:genetics; D005260:Female; D015684:Gene Products, pol / Q000737:chemistry / Q000235:genetics*; D014644:Genetic Variation; D006515:Hepatitis B virus / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000188:drug therapy / Q000821:virology; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007214:Indonesia; D008297:Male; D008875:Middle Aged; D000072417:Protein Domains; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA; D055815:Young Adult
chemical_list: D000998:Antiviral Agents; D015684:Gene Products, pol; C066027:P protein, Hepatitis B virus; D012194:RNA-Directed DNA Polymerase
keywords: Drug resistance; Hepatitis B virus; Indonesia; Reverse transcriptase domain; Ultra-deep sequencing