Sampling algorithm for geno- and subtyping of hepatitis B virus using panel of monoclonal antibodies in enzyme immunoassay

Abstract

Introduction. Previously, a domestic method for determining hepatitis B virus (HBV) genotypes and HBV surface antigen (HBsAg) subtypes in HBsAgpositive blood serum samples with a Monoclonal Antibody Panel (MAB) by enzyme immunoassay (ELISA) was described. In routine laboratory practice, HBsAg detection results are obtained in units of optical density (OD).Purpose. To describe the sampling algorithm for ELISA geno- and subtyping of HBV with the MAB Panel.Materials and methods. We studied 40 blood serum samples with positive results of HBsAg determination. HBV genotypes and HBsAg subtypes were determined using four different MAB conjugates with horseradish peroxidase using the ELISA method described previously.Results. In samples with OD less than 2.0 o. u. and confirmation of HBsAg after a single confirmatory study (n = 19) HBV genotypes/ HBsAg subtypes not established. In samples with an OD above 2.0 o. u. and verification of the presence of the antigen in the standard mode of registration (n = 15), immunoenzymatic geno- and subtyping of HBV was effective in 27 % of cases (4/15); for samples with HBsAg verification carried out in an auxiliary measurement (n = 6) the effectiveness of the technique was 100 %. Using the MAB panel in ELISA, the characteristics of HBV in 10 samples with the presence of HBsAg were studied: HBV genotype D (n = 10), subtypes ayw2 (n = 7), ayw3 (n = 3).Conclusions. For the most effective application of the HBV geno- and subtyping technique using the MAB Panel in ELISA, HBsAg+ samples with OD signals of more than 2.0 o. u. in screening and verification of the presence of an antigen in the auxiliary mode of registration of results should be used.