[Line immunoassay for detection of IgG antibodies to hepatitis E virus (Hepeviridae, Orthohepevirus, Orthohepevirus A)]

Vopr Virusol. 2020 Jul 21;65(3):132-142. doi: 10.36233/0507-4088-2020-65-3-132-142.
[Article in Russian]

Abstract

Introduction: The diagnostic efficacy of methods for hepatitis E serodiagnostic varies over a wide range; therefore, the combined use of tests of various formats is recommended. The aim of the research was to develop a test system for the detection of IgG antibodies to hepatitis E virus (HEV) in human serum by linear immunoassay (LIA).

Material and methods: Serum samples from patients with hepatitis and healthy individuals were tested using commercial enzyme-linked immunosorbent assay systems for the presence of IgG antibodies to viral agents causing hepatitis and other infections associated with liver pathology. Recombinant antigens ORF2 and ORF3 of HEV genotypes 1 and 3 were used. The "RecomLine HEV IgG/IgM" reagent kit (Mikrogen GmbH, Germany) was used as a comparison test system.

Results: The first Russian diagnostic kit "Blot-HEV", designed to detect IgG antibodies to individual HEV proteins in human serum using LIA, was developed. The antigenic base is represented by strips of a nitrocellulose membrane with immobilized recombinant antigens ORF2 (aa 406-660) and ORF3 (aa 1-113) of HEV genotypes 1 and 3, and control antigens in the form of discrete lines. The conjugate was mouse monoclonal antibodies to human class G immunoglobulins labeled with horseradish peroxidase. The chromogen solution contained the 3,3',5,5'-tetramethylbenzidine. A visual and digital recording of results was provided. The analytical sensitivity of the test kit was 0.625 IU/ml for ORF2 antigens and 2.5 IU/ml for ORF3 antigens. The absence of the influence of endogenous interfering substances on the results of the analysis and the absence of cross-reactions with antibodies to hepatitis pathogens of the other etiologies had been shown. The sensitivity of the test system compared to the "RecomLine HEV IgG/IgM" kit was 92%, specificity 97%. Shelf life in condition of storage was determined to be 12 months.

Conclusions: The developed test can be used to confirm the results of ELISA in laboratory diagnosis of hepatitis E.

MeSH terms

  • Adult
  • Female
  • Hepatitis Antibodies / blood
  • Hepatitis E / blood*
  • Hepatitis E / virology
  • Hepatitis E virus / genetics
  • Hepatitis E virus / isolation & purification*
  • Hepatitis E virus / pathogenicity
  • Humans
  • Immunoassay*
  • Immunoglobulin G / blood*
  • Immunoglobulin G / isolation & purification
  • Male
  • Russia / epidemiology

Substances

  • Hepatitis Antibodies
  • Immunoglobulin G