Hepatitis B Virus Genotypes in Patients with Hepatitis D as Determined by the Panel of Their Own Development of Monoclonal Antibodies

Direct genotyping of hepatitis B virus (HBV) in samples from patients with hepatitis delta can be impossible due to an undetectable concentration of HBV DNA. A sufficient amount of surface HBV protein (HBsAg) in such samples makes it possible to determine HBV genotype using enzyme-linked immunosorbent assay (ELISA) of this antigen with a panel of monoclonal antibodies (MABs). The purpose of this paper is to compare the results of HBV genotyping using an in-house MAB panel with the results of molecular analysis of samples from patients with chronic hepatitis B (CHB) and the determination of HBV genotypes in samples from patients with hepatitis delta. A total of 122 serum samples from CHB patients from Yakutia and 211 serum samples from hepatitis delta patients from Yakutia (12 samples) and Tuva (199 samples) were collected. HBV serotypes/genotypes were determined by ELISA using developed reagents. The molecular methods included the isolation of HBV DNA, amplification of the S gene region (713 nt), and phylogenetic analysis of the nucleotide sequences. In a group of samples from CHB patients positive for HBV DNA (86 samples), 95% (82/86) valid results were obtained using our MAB kit. The following genotypes were identified: A, 32 (39%); C, 3 (4%); and D, 47 (57%). The results of HBV genotyping using two methods were identical for 81/82 (99%) samples. HBV genotyping using developed reagents provided 96.2% (203/211) of valid results in samples from patients with hepatitis delta compared to 3.8% of such results obtained using the standard molecular technic ( p < 0.001). The following HBV genotypes were identified: A, 17 samples (8.4%), and D, 186 samples (91.6%). The developed test with MAB panel allows one to reliably determine HBV genotype and has advantages over standard molecular methods for HBV genotyping in patients with hepatitis delta.