Development and evaluation of an enzyme immunoassay for diagnosis of acute viral hepatitis E
Year: 2025
Type: article
Abstract: Objective . To develop a national high-sensitivity and marked specifity enzyme immunoassay test system for determining class М immunoglobulins to the hepatitis E virus (HEV) in human blood serum “ELISA-anti-HEV IgМ“. Materials and methods . The base for this research is City Clinical Hospital of Infectious Diseases in Minsk. Qualitative determination of anti-HEV IgМ in human blood serum samples was carried out by indirect enzyme immunoassay using reference kits – “ELISA-ANTI-HEV-IgМ“, Vecto hep E-IgМ. 96-well polystyrol strip plates were used to design a national test system for the qualitative detection of anti-HЕV-IgМ; recombinant antigens – analogues of the protein ORF2 and ORF3 HЕV of the genotype 3; antibodies against human IgM conjugated with horseradish peroxidase; post-closing solution; serum dilution solution and conjugate dilution solution. Results . Optimal concentrations for sorption of ORF2 and ORF3 proteins were determined, amounting to 2 mg/ml and 1 mg/ml. The incubation time with TMB is set to 20 minutes. Diagnostic sensitivity of the test system was no less than 99%, and diagnostic specificity was no less than 99%. The index of in-series reproducibility was 92,3%, inter-series reproducibility was 86,1%. Technical specifications TU BY 100185093.093-2023 were developed and registered. Direction for use of the reagent kit of the test system “ELISA-anti-HEV-IgM human“ for determining markers of acute viral hepatitis E in human blood serum by enzyme immunoassay was approved. Conclusion . The first national test system “ELISA-anti-HЕV-IgM” for the determination of class M immunoglobulins against hepatitis E virus in human blood serum by indirect enzyme immunoassay was developed. Laboratory and technical tests of the experimental sample of the test system were carried out, shown its high diagnostic sensitivity, specificity and reproducibility. The regulatory, technical and operational documentation for the test system was approved.
Source: Health and Ecology Issues
Language: English
Subfield: Hepatology
Field: Medicine
Domain: Health Sciences
Open Access status: hybrid
OpenAlex ID: W7128242560