IDENTIFICATION OF X GENE MUTATIONS, GENOTYPE AND SUBGENOTYPE OF HEPATITIS B VIRUS (HBV)
Abstract: Infection of
hepatitis B virus (HBV) is a global health problem, including in Indonesia.
There are currently an estimated 2 billion people worldwide are infected with
HBV, about 75% of them are in Asia and 350 million of them will develop into
chronic hepatitis B. Factors that influence the development of chronic
hepatitis B into liver carcinoma include x gene mutation, HBV genotype and
subgenotype. This research aims to identify x gene mutation, genotype and
subgenotype of HBV infecting hepatitis B patients in Manado associated with the
onset of liver carcinoma. HBV DNAs were isolated from blood samples of 30
hepatitis B patients. X gen was amplified using nested PCR with pre-designed
primer pairs. Amplified DNA fragments were electrophoresed in 1.5% agarose and
visualized under UV. DNA fragments were then separated and purified using
Qiagen column, then sequenced to determine their nucleotide sequences of x
gene. Amino acid of x protein were deduced from nucleotide sequence of x gene
and used as basic to determine HBV genotype and subgenotype. X protein was
aligned with those similar protein with the same subgenotype retrieved from
GenBank to determine if there was a mutation at amino acid. The mutated x protein were compared with
other mutation found in x protein in other literatures associated with the
onset of liver carcinoma. Genotype and subgenotype of HBV isolated from blood
samples of 10 patients was detected and showed that five patients were infected
with B genotype HBV (2 patients were infected with B2 subgenotype, 2 patients
with B3 subgenotype and 1 patient with B9 subgenotype). The rest of 5 patients
were infected with C genotype HBV (1 patient with C1 subgenotype, 2 patients
with C2 subgenotype, and 2 patients with
C5 subgenotype). The mutation in x protein is related significantly to the
clinical severity of the liver and hepatocellular carcinoma (HCC), ie V5L in subgenotype C2,
and I127T and H94Y in subgenotype C5.
Author: Fatimawali, Billy
Kepel, Sitti Romlah, Catur Riany, Debbie Retnoningrum
Journal Code: jpfarmasigg140039
