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Background: Hepatitis B virus (HBV) induces both acute and chronic hepatitis in humans. Although a safe and effective vaccine exists, HBV infection continues to pose a major global health challenge. Chronic HBV affects approximately 240–300 million individuals worldwide. Annually, about 600,000 fatalities occur due to liver diseases associated with HBV. Aims: This study aims to detect prevalent genotypes in cases of hepatitis B attending a tertiary care hospital. Settings and Design: A hospital-based prospective study was conducted in the viral research and diagnostic laboratory from January 1, 2023, to June 30, 2024. Materials and Methods: A 5 ml blood sample was collected from each patient and tested for hepatitis B surface antigen (HBsAg) using chemi enzyme-linked immunosorbent assay on the Abbott ARCHITECT i1000SR (Abbott Diagnostics, Chicago, IL, USA) platform. A total of 76,896 patients screened, 759 tested positive. Among them, 588 patients had detectable viral load using the Truenat ® assay. Deoxyribonucleic acid was extracted from 100 samples with high viral loads using QIAamp ® kit. Real-time polymerase chain reaction was used for genotype analysis using Sacace™ HBV Genotype kit. Results: A total of 76,896 patients were screened for HBsAg, revealing 759 positive cases (0.98% seroprevalence). Genotype D was found to be more prevalent, comprising 68% compared to 32% for Genotype A, with the highest number in 20–29 years of age group (29%), where 69% were Genotype D. The youngest age group (10–19) showed a contrasting trend, with 75% being Genotype A. Conclusions: Genotype D is prevalent in our region. To reduce HBV transmission, vaccination and public health interventions should target predominant genotypes.
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