Abstract
Background: Hepatitis B virus (HBV) infection remains a major global health challenge, especially among high-risk groups such as hemodialysis (HD) patients. Aim: This study investigated the prevalence of sero-molecular markers and the genetic diversity of HBV in 160 Palestinian HD patients. Blood samples were tested for HBV serological markers (HBsAg, anti-HBc, and anti-HBs) and screened using nested PCR. Whole genome sequencing was conducted on PCR-positive samples to identify HBV genotypes and subgenotypes. Results: The overall HBV prevalence among HD patients was 3.75%, comprising 1.9% with overt infection (HBsAg +ve) and 1.9% with occult HBV infection (OBI). HCV was detected in 1.9% of patients. Evidence of past exposure (anti-HBc positive) was observed in 20% of patients, and 45% showed serological immunity with anti-HBs levels ≥ 10 IU/mL. Although the values of the genetic diversity estimators such as K, S, η, and π were approximately as twice as those for the S-region, the S-region produced a more reasonable phylogenetic tree and haplotype networking but under the condition of accurate sequencing and adequate number of investigated sequences. Phylogenetic trees and haplotype networking of the WGS and S-region revealed a clustering pattern based on genotypes and subgenotypes with two Palestinian WGS clustering in Subgenotype D1, while the other two in Subgenotype D3. Genetic diversity analysis revealed high haplotype diversity (Hd) (0.98-1.00) with high h:n ratio (0.9-1.00) and low nucleotide diversity (π) (0.007-0.027) indicating slight variation between any two given sequences. This is explained by purifying selection, recent population expansion, or constrained evolution as neutrality test values such as Tajima's D were negative (-0.5 to -1.86). Conclusion: HBV infection remains prevalent among HD patients, including both overt and occult forms. Genotype D, specifically Subgenotypes D1 and D3, predominates in the study population. The HBV S-region is a sufficient surrogate for population genetics investigations.