A deep-sequencing method detects drug-resistant mutations in the hepatitis B virus in Indonesians.
OBJECTIVE
The long-term administration of a nucleos(t)ide analogue (NA) for the treatment of chronic hepatitis B may encourage the emergence of viral mutations associated with drug resistance. Minor populations of viruses may exist before treatment, but are difficult to detect because of technological limitations. Identifying minor viral quasispecies should be useful in the clinical management of hepatitis B virus (HBV) infection.


METHODS
Six treatment-naïve Indonesian patients with chronic HBV infection participated in this study. The polymerase region of the HBV genome, including regions with known drug-resistant mutations, was subjected to capillary sequencing and MiSeq sequencing (Illumina). Mutations were analyzed with Genomics Workbench software version 6.0.1 (CLC bio).


RESULTS
The mean mapping reads for the six samples was 745,654, and the mean number of amplified fragments ranged from 17,926 to 25,336 DNA reads. Several known drug-resistant mutations in the reverse transcriptase region were identified in all patients, although the frequencies were low (0.12-1.06%). The proportions of the total number of reads containing mutations I169L/M, S202R, M204I/L or N236S were >1.0%.


CONCLUSIONS
Several known NA-resistant mutations were detected in treatment-naïve patients in Indonesia using deep sequencing. Careful management of such patients is essential to prevent drug-resistant mutations from spreading to other patients.
Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan.
Dewiyani Indah
DI
Widasari
Yoshihiko
Y
Yano
Didik Setyo
DS
Heriyanto
Takako
T
Utsumi
Laura Navika
LN
Yamani
Hanggoro Tri
HT
Rinonce
Widya
W
Wasityastuti
Maria Inge
MI
Lusida
Soetjipto
Rina
R
Okada
Yoshiki
Y
Murakami
Toshihito
T
Tanahashi
Takeshi
T
Azuma
Yoshitake
Y
Hayashi
D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology*; D024882:Drug Resistance, Viral / Q000235:genetics*; D005838:Genotype; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000821:virology; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007214:Indonesia; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*; D055815:Young Adult
D000998:Antiviral Agents; D012194:RNA-Directed DNA Polymerase