In-house primer panel driven resource-efficient whole-genome sequencing of hepatitis B virus

Area of Science:

• Virology
• Genomics
• Epidemiology

Background:

• Hepatitis B virus (HBV) genetic variability impacts disease progression and treatment.
• Limited whole-genome sequencing (WGS) data exists for HBV in Central India.
• Standardized commercial assays for HBV WGS are lacking in the region.

Purpose of the Study:

• To develop an in-house, primer-based HBV WGS workflow.
• To characterize the molecular epidemiology and mutational profile of HBV in Chhattisgarh, India.
• To establish a cost-effective method for molecular surveillance in resource-limited settings.

Main Methods:

• 102 HBV-positive clinical samples were analyzed.
• Whole-genome sequencing (WGS) was performed using in-house designed overlapping primers.
• Sequencing quality, genome coverage, phylogenetic relationships, and mutational patterns were evaluated.

Main Results:

• High-quality WGS was achieved with >98.7% genome coverage.
• HBV genotype D was exclusively predominant, with sub-genotype D3 being most common.
• Frequent mutations were identified in the surface, transmembrane, and core regions, including the precore mutation C1858T.

Conclusions:

• An effective and cost-efficient in-house HBV WGS approach was established.
• A minimum viral load of 3.11 log₁₀ IU/mL is suggested for adequate genome coverage.
• The predominance of genotype D and identified mutations emphasize the need for genomic monitoring for enhanced HBV control.