Hepatitis B Virus Genotypes in Patients with Hepatitis D
as Determined by the Panel of Their Own Development
of Monoclonal Antibodies
https://orcid.org/0000-0001-9527-8870L. V. Bezuglovaa, * (), https://orcid.org/0000-0002-2656-3667O. V. Isaevab, c (),
https://orcid.org/0000-0002-6013-7768A. A. Karlsenb, c (), https://orcid.org/0000-0001-6029-1864L. Y. Ilchenkoc, d (),
https://orcid.org/0000-0002-0103-4750S. S. Sleptsovae (), https://orcid.org/0000-0002-2116-9839A. A. Saryglarf (),
https://orcid.org/0000-0003-1503-8426V. A. Poryvaevaa (), https://orcid.org/0000-0001-8801-1297Ya. D. Mosinaa (),
https://orcid.org/0000-0001-9887-6067O. A. Agafonovaa (), https://orcid.org/0000-0003-2884-729XA. K. Mogilnykha (),
https://orcid.org/0000-0002-3599-117XK. K. Kyuregyanb, c (), https://orcid.org/0000-0002-6636-6801M. I. Mikhailovb, c (),
https://orcid.org/0000-0002-7786-2464S. V. Netesovg (), and https://orcid.org/0000-0002-1953-5123I. G. Netesovaa ()
a AO Vector-Best, Koltsovo, 630559 Russia
b Russian Medical Academy of Continuous Professional Education, Moscow, 125993 Russia
c Mechnikov Research Institute for Vaccines and Sera, Moscow, 105064 Russia
d Pirogov Russian National Research Medical University, Moscow, 117997 Russia
e North Eastern Federal University, Medical Institute, Yakutsk, 677000 Russia
f Kyzyl Infectious Hospital, Kyzyl, 667005 Russia
g Novosibirsk State University, Novosibirsk, 630090 Russia
Correspondence to: *e-mail: bezuglova@vector-best.ru
Received 12 August, 2020
Abstract—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.
Keywords: HBsAg, hepatitis B virus, genotype, monoclonal antibodies, enzyme-linked immunosorbent assay, hepatitis D
DOI: 10.3103/S0891416822020033