Molecular Characterization Of Hepatitis B Virus In Rural And Semi-Urban Areas In The Central Region Of Ghana

ABSTRACT Hepatitis B virus (HBV) infection is a major cause of liver inflammation accounting for 887,000 deaths annually. The introduction of HBV vaccines has significantly reduced new cases of the infection, but there are still 65 million people living with the virus in Sub-Saharan Africa. Currently, 10 genotypes (A-J) of HBV have been classified. Several studies have shown that the different HBV genotypes differently influence clinical presentation, progression of liver diseases and response to treatment. In Ghana, studies on HBV circulating genotypes and their implications on epidemiology is limited, however few available data have been obtained mostly in the urban areas of the Ashanti and Greater Accra regions. This study is aimed at characterizing HBV genotypes in patients attending three district hospitals in the Central Region of Ghana (Agona-West, Effutu Municipal and Gomoa-West district hospitals). Questionnaires were administered to the patients before sample collection which included their gender, age, educational levels and risk factors. There was no association between socio-demographic factors and HBV infectivity and transmissibility, except educational levels. HBV DNA was extracted from the blood of 173 HBsAg seropositive patients using QIAamp DNA Blood mini kit; 70 (40%) samples from the Gomoa-West district, 50 (29%) samples from the Effutu Municipal and 53 (31%) from the Agona-West district. The extracted DNA was confirmed by amplifying the S region of the HBV genome using conventional PCR. A total of 115 (66.5%) patients were found to be PCR positive, with 58 (33.5%) negative for HBV DNA. The HBV was genotyped by nested-multiplex PCR using type-specific primers that amplifies the preS/S region of the HBV genome. To confirm the specificity of the nested-multiplex PCR assay, 31 of the samples obtained from the S gene amplification were sent for Sanger sequencing in one direction. The nucleotide sequences obtained iii were submitted into the HBV HepSeq genotyping software to confirm the genotypes. The genotype results obtained from both the software and the nested -multiplex PCR assay included; 67.1% genotype E which was the most dominant genotype, followed by genotypes A, D and G with 1.7%, 0.6% and 0.6%, respectively. The phylogenetic analysis of the sequenced isolates from the three districts clustered together with the reference strains. The characterization of the HBV genotypes will help understand the transmission dynamics of the virus in the rural and semi-urban setting.