Fatimawali ., Billy Kepel, Sitti Romlah, Catur Riany, Debbie Retnoningrum


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.

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Barbini L., Tadey L., Fernandez S., Bouzas B., Campos R. 2012. Molecular characterization of hepatitis B virus X gene in Chronic hepatitis b patients, Virology Journal, open acces, 9:131

Baumert TF., Thimme R., Weizsäcker FV. 2007, Pathogenesis of hepatitis B virus infection, World J Gastroenterol 2007 January 7; 13(1): 82-90

Gunawan VA. 2012. Genotype and Subgenotype Determination of hepatitis B Virus from Patiens by Using Gene Sequence as a Basis for Counseling, Thesis, Sekolah Farmasi ITB Bandung.

Hann HWL. 2007. Hepatitis B Virus Screening and Counseling Strategies for General Practitioners, John Hopkins Advanced Studies in Medicine, 7(15), 476-481.

Kao JH., Wu NH., Chen PJ., Lai MY., Chen, DS., 2000, Hepatitis B Genotypes and The response to Interferon Therapy, J. Hepatol, Vol. 33, Issue 6, Pages 998-1002.

Kao JH., Liu CJ., Chen DS. 2002. Hepatitis B viral genotypes and lamivudine resistance. J Hepatol; 36:303-304.

Kim BJ. 2014. Hepatitis B Virus Mutations Related to Liver Desease Progression of Korean Patients, World Journal of Gastroenterology.

Kim HJ., Park JH., Jee Y., Lee SA., Kim H., et al., 2008. Hepatitis B Virus X Mutations Occurring Naturally Associated With Clinical Severity of Liver Disease Among Korean Patients With Chronic Genotype C Infection, J Med Virol 80:1337–1343

Lee JH., Han KH., Lee JM., Park JH., Kim, HS., 2011, Impact of Hepatitis B Virus (HBV) X Gene Mutations on Hepatocellular Carcinoma Development in Chronic HBV Infection, Clin Vaccine Immunol., 18(6): 914–921

Liu, S.H., Zhang,C. Gu, J. Yin,Y., He, Xie j., and Cao G. 2008, Associations Between Hepatitis B Virus Mutations and the Risk of Hepatocellular Carcinoma: A Meta-Analysis, J. Natl. Canc. Inst., 101(15), 1066-1082.

Lupberger J., Hildt E., 2007, Hepatitis B virus-induced oncogenesis, World J Gastroenterol. 13(1): 74-81

Mulyanto, Depamede SN., Kiely S., Tsuda F., Ichiyama K., Takahashi M., Okamoto H. 2009. A Nationwide Molecular Epidemiological Study on Hepatitis B Virus in Indonesia: Identification of two novel subgenotypes, B8 and C7, Arch Virol, DOI 10.1007/s00705-009-0406-9.

Rini KR. 2013. Analisis Mutasi Gen x Virus Hepatitis B pada pasien dengan Metode Nested PCR sebagai dasar Konseling, Skripsi, Sekolah Farmasi ITB Bandung.

Wai CT., Chu CJ., Hussain M., Lok ASF. 2007. HBV Genotype B Is Associated With Better Response to Interferon Therapy in HBeAg(+) Chronic Hepatitis Than Genotype C, J. Hepatology; 36; 1425-1430.

Wang X., Chen Y., Yu D., Zhang W., Qiu C., Xiang G., Dai W., Wu, S. 201., HBV Subgenotype C2 Infection, A1762T/ G1764A Mutations May Contribute To Hepatocellular Carcinoma with Cirrhosis in Southeast China, Iranian J. Publ Health, 41(11), 10-18.

Yang HI., Yeh SH., Chen PJ., Iloeje UH., Jen C.L., et al., 2008, Associations Between Hepatitis B Virus Genotype and Mutants and the Risk of Hepatocellular, J Natl Cancer Inst , 2008;100: 1134 – 1143.



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