The aim of this study was to assess the prevalence of the occult hepatitis B infection among blood donors in St. Petersburg, as well as to characterize the identified virus isolates. The study material was represented by 2800 blood plasma samples collected in 2019 from blood donors living in St. Petersburg. The ELISA examination for HBV markers occurrence involved HBsAg, anti-HBs IgG, anti-HBcore IgG. HBV DNA was detected by nested PCR with real-time hybridization-fluorescence detection on three targets, which makes it possible to determine virus DNA at low viral load, including HBsAg-negative chronic hepatitis B. Hepatitis B serological markers were detected in 69.43 % of those surveyed, HBsAg was found in 0.43% of individuals, and all of them donated blood for the first time. A significant excess of the anti-HBcore IgG antibodies occurrence among primary donors (15.1%) compared with repeated/regular donors (7.48%) was shown. The occurrence of virus DNA in the group was 3.14%, including 2.71% of cases representing HBsAg-negative CHB. Based on phylogenetic analysis of 88 isolates, HBV subgenotypes were determined in the following ratios: D1 and D2, 40.91% each, D3 and A2, 9.09% each. When determining the serological subtype of the detected isolates, the serotype ayw3 (52.27%) prevailed compared to ayw2 (46.59%) and adw2 (10.23%). Drug resistance mutations, including compensatory ones, were detected in six examined patients (6.82%). In all genotype D isolates, multiple amino acid substitutions were identified in the RT, SHB, MHB, LHB, and Core regions; mutations in the preCore region were detected in 21.59% of the samples. In the MHR of the HBV genotype D genome, twenty-six positions were identified in which amino acid substitutions occurred, and all isolates showed modifications at positions 113, 114, 131, 134, 159, 161, 168, in 76 - in position 122, in 68 - in position 127, in 36 - in position 118, in 24 - in position 128. In HBV A2 isolates, mutations T113S, S143T, Y161F were identified. Nine isolates in the preCore region showed a polymorphism including a stop codon: W28*W; the W28S substitution was shown in the same position in five isolates, and the W28*S variant was identified in one more sample.
The high incidence of HBsAg-negative CHB cases among blood donors, as well as the predominance HBV isolates that simultaneously carry mutations that lead to diagnostic failure of HBsAg tests and prophylactic failure of immunoglobulin or vaccines and virus reactivation, mutations that contribute to the progression of the disease, obviously pose a threat health and needs further study.