Methodological basics for differential detection of EBV1/EBV2 and HHV6A/HHV6B

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Abstract

Among a whole variety of EBV- and HHV6-linked diseases only infectious mononucleosis is subject to official statistical reporting in Russia that substantially complicates objective assessment of etiological structure, incidence rate, characteristics of developing epidemic process. Currently, the data on the genetic EBV heterogeneity, even at the level of the main types (EBV1 and EBV2), as well as HHV6A and HHV6B, prevalence and clinical importance are mainly limited to foreign research publications. Few publications assessing this issue are available in Russian scientific papers. At the same time, examining circulation of virus genetic types (variants) and use of such data in implementing epidemiological surveillance after some other infections have been commonly practiced. One of the key issues is the level of developed laboratory support for molecular genetic monitoring. The goal of the study was to improve methodological basics for differential detection of HHV6A/B and the major EBV types. There were used samples of peripheral blood leukocytes collected from children aged 1–15 years with acute (n = 50) and asymptomatic infectious mononucleosis (n = 29). The detection and quantification of EBV and HHV6 DNA was performed by using real-time PCR. For differential determination of EBV1/EBV2 and HHV6A/HHV6B, an optimized one-round PCR with electrophoretic agarose gel detection amplification products was used. The data from our own study showed that frequency of detected EBV and HHV6 DNA in acute infectious mononucleosis patients comprised 74 and 72% compared to control group reaching 35 and 74%, respectively. It was found that among the examined children of Nizhny Novgorod Region, EBV1 and HHV6B prevailed in the viral population that agrees with existing insights about their geographical distribution in the adjacent territories. EBV2 was found in a single sample only in the control group. HHV6A was not detected in any of the groups. The methodological approach optimized in this study allows to separately detect HHV6A/HHV6B and the main EBV types according to a unified laboratory protocol, whereas combining it with additional stage of DNA enrichment increases the diagnostic sensitivity of PCR analysis, minimizes proportion of discordant and false negative results. Such an integrated approach can be applied for diagnostic, epidemiological and research purposes.

About the authors

M. I. Popkova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology

Author for correspondence.
Email: popmarig@mail.ru
ORCID iD: 0000-0001-5864-5862

Mariia I. Popkova - PhD (Мedicine), Leading Researcher, Laboratory of Molecular Biology and Biotechnology, Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology.

603950, Nizhny Novgorod, Malaya Yamskaya str., 71.

Phone: +7 (831) 469-79-46. Fax: +7 (831) 469-79-20.

SPIN: 4485-2459

Russian Federation

O. V. Utkin

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology

Email: utkino2004@mail.ru
ORCID iD: 0000-0002-7571-525X

PhD (Biology), Leading Researcher, Laboratory of Molecular Biology and Biotechnology, Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology.

Nizhny Novgorod.

SPIN: 1174-6500

Russian Federation

E. A. Soboleva

Privolzhsky Research Medical University

Email: Fullofcarrot@pimunn.ru
ORCID iD: 0000-0003-3591-9618

Ifectologist, Assistant Professor, Department of Infectious Diseases, Privolzhsky Research Medical University.

Nizhny Novgorod.

Russian Federation

N. A. Sakharnov

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology

Email: saharnov@nniiem.ru
ORCID iD: 0000-0003-3965-2033

Researcher, Laboratory of Molecular Biology and Biotechnology, Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology.

Nizhny Novgorod.

SPIN:8457-3501

Russian Federation

D. A. Bryzgalova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology

Email: moskvinadara7@gmail.com
ORCID iD: 0000-0002-6663-8440

Junior Researcher, Laboratory of Molecular Biology and Biotechnology, Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology.

Nizhny Novgorod.

SPIN: 5626-3491

Russian Federation

A. O. Senatskaia

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology

Email: seanna@bk.ru
ORCID iD: 0000-0003-2024-4283

Junior Researcher, Laboratory of Molecular Biology and Biotechnology, Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology.

Nizhny Novgorod.

SPIN: 6891-2140

Russian Federation

E. A. Kulova

Privolzhsky Research Medical University

Email: dr_kulova@mail.ru
ORCID iD: 0000-0002-5207-1164

PhD (Medicine), Assistant Professor, Department of Infectious Diseases, Privolzhsky Research Medical University.

Nizhny Novgorod.

SPIN: 4331-6958

Russian Federation

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Copyright (c) 2021 Popkova M.I., Utkin O.V., Soboleva E.A., Sakharnov N.A., Bryzgalova D.A., Senatskaia A.O., Kulova E.A.

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