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

G. I. Alatortseva; Алаторцева Г. И.; V. V. Dotsenko; Доценко В. В.; L. N. Nesterenko; Нестеренко Л. Н.; L. N. Luhverchik; Лухверчик Л. Н.; V. Yu. Kabargina; Кабаргина В. Ю.; I. I. Amiantova; Амиантова И. И.; M. V. Zhukina; Жукина М. В.; S. V. Zhavoronok; Жаворонок С. В.; Z. S. Nurmatov; Нурматов З. Ш.; A. Z. Nurmatov; Нурматов А. З.; V. V. Zverev; Зверев В. В.

doi:10.36233/0507-4088-2020-65-3-132-142

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

Authors: Alatortseva G.I.¹, Dotsenko V.V.¹, Nesterenko L.N.¹, Luhverchik L.N.¹, Kabargina V.Y.¹, Amiantova I.I.¹, Zhukina M.V.¹, Zhavoronok S.V.², Nurmatov Z.S.³, Nurmatov A.Z.³, Zverev V.V.¹
Affiliations:
1. I.I. Mechnikov Research Institute of Vaccines and Sera
2. Belarusian State Medical University
3. Scientific Production Association “Preventive Medicine” Ministry of Health of the Kyrgyz Republic
Issue: Vol 65, No 3 (2020)
Pages: 132-142
Section: ORIGINAL RESEARCH
URL: https://journals.rcsi.science/0507-4088/article/view/118126
DOI: https://doi.org/10.36233/0507-4088-2020-65-3-132-142
ID: 118126

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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.

Keywords

hepatitis E, hepatitis E virus, serodiagnosis, line immunoassay, immunoblotting

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Bishkek, 720005

Kyrgyzstan

V. V. Zverev

I.I. Mechnikov Research Institute of Vaccines and Sera

Email: fake@neicon.ru
ORCID iD: 0000-0002-0017-1892

Vitaliy V. Zverev

Moscow, 105064

Russian Federation

References

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