biopreparatБИОпрепараты. Профилактика, диагностика, лечениеBiological Products. Prevention, Diagnosis, Treatment2221-996X2619-1156Scientific Centre for Expert Evaluation of Medicinal Products10.30895/2221-996X-2026-750biopreparat-750Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESГенетическая характеристика производственных штаммов вирусов кори, краснухи и паротита методом высокопроизводительного секвенированияGenetic characterization of production strains of measles, rubella, and mumps viruses by high-throughput sequencinghttps://orcid.org/0000-0001-8674-6097ЖелтухинаА. И.ZheltukhinaA. I.

Желтухина Алена Игоревна

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Alyona I. Zheltukhina

15/17 Professor Popov St., St. Petersburg 197376

alena.zheltuhina@influenza.spb.ruhttps://orcid.org/0000-0003-1733-1255КомиссаровА. Б.KomissarovA. B.

Комиссаров Андрей Борисович 

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Andrey B. Komissarov

15/17 Professor Popov St., St. Petersburg 197376

andrey.komissarov@influenza.spb.ruhttps://orcid.org/0000-0003-3558-3261ФадеевА. В.FadeevA. V.

Фадеев Артем Викторович 

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Artem V. Fadeev

15/17 Professor Popov St., St. Petersburg 197376

artem.fadeev@influenza.spb.ruhttps://orcid.org/0000-0002-1050-5817ЁлшинН. Д.YolshinN. D.

Ёлшин Никита Дмитриевич 

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Nikita D. Yolshin

15/17 Professor Popov St., St. Petersburg 197376

nikita.yolshin@gmail.comhttps://orcid.org/0000-0002-1499-9957ПисареваМ. М.PisarevaM. M.

Писарева Мария Михайловна, канд. биол. наук 

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Maria M. Pisareva, Cand. Sci. (Biol.)

15/17 Professor Popov St., St. Petersburg 197376

maria.pisareva@influenza.spb.ruhttps://orcid.org/0000-0002-9970-3325ЕдерВ. А.EderV. A.

Едер Вероника Анатольевна, канд. биол. наук 

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Veronika A. Eder, Cand. Sci. (Biol.)

15/17 Professor Popov St., St. Petersburg 197376

veronika.eder@influenza.spb.ruhttps://orcid.org/0000-0003-2512-2517КоржановаМ.KorzhanovaM.

Коржанова Маргарита 

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Margarita Korzhanova

15/17 Professor Popov St., St. Petersburg 197376

marory.kmm@gmail.comhttps://orcid.org/0000-0002-4532-6210КсенафонтовА. Д.KsenafontovA. D.

Ксенафонтов Андрей Дмитриевич

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Andrey D. Ksenafontov

15/17 Professor Popov St., St. Petersburg 197376

ksenandrey@yandex.ruhttps://orcid.org/0000-0002-5961-1856ПередерийА. А.PerederiyA. A.

Передерий Александр Александрович

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Alexander A. Perederiy

15/17 Professor Popov St., St. Petersburg 197376

gilagalex@gmail.comhttps://orcid.org/0000-0002-8214-7828ОхлопковаО. В.OhlopkovaO. V.

Охлопкова Олеся Викторовна, канд. биол. наук

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Olesia V. Ohlopkova, Cand. Sci. (Biol.)

15/17 Professor Popov St., St. Petersburg 197376

ohlopkova.lesia@yandex.ruhttps://orcid.org/0009-0004-2369-3767СаушкинаЛ. А.SaushkinaL. A.

Саушкина Людмила Александровна

2-й Волконский переулок, д. 10, Москва, 127473

Lyudmila A. Saushkina

10 2nd Volkonsky Lane, Moscow 127473

hrisa07@rambler.ruhttps://orcid.org/0000-0002-9229-3677ШведоваЕ. В.ShvedovaE. V.

Шведова Евгения Владимировна, канд. биол. наук

2-й Волконский переулок, д. 10, Москва, 127473

Evgenia V. Shvedova, Cand. Sci. (Biol.)

10 2nd Volkonsky Lane, Moscow 127473

89057336653@mail.ruhttps://orcid.org/0000-0002-5621-8266ГришинаЕ. А.GrishinaE. A.

Гришина Елена Анатольевна, д-р биол. наук, проф.

2-й Волконский переулок, д. 10, Москва, 127473

Elena A. Grishina, Dr. Sci. (Biol.), Prof.

10 2nd Volkonsky Lane, Moscow 127473

Gelana2010@yandex.ruhttps://orcid.org/0000-0003-3643-7354ЛиозновД. А.LioznovD. A.

Лиознов Дмитрий Анатольевич, д-р мед. наук, проф.

ул. Профессора Попова, д. 15/17, Санкт-Петербург, 197376

Dmitry A. Lioznov, Dr. Sci. (Med.), Prof.

15/17 Professor Popov St., St. Petersburg 197376

office@influenza.spb.ruФедеральное государственное бюджетное учреждение «Научно-исследовательский институт гриппа имени А.А. Смородинцева» Министерства здравоохранения Российской ФедерацииРоссияA.A. Smorodintsev Research Institute of InfluenzaRussian FederationАкционерное общество «Научно-производственное объединение по медицинским иммунобиологическим препаратам «Микроген»РоссияScientific and Production Association MicrogenRussian Federation20261208202600Принято в печать (Online First)750Copyright © Желтухина А.И., Комиссаров А.Б., Фадеев А.В., Ёлшин Н.Д., Писарева М.М., Едер В.А., Коржанова М., Ксенафонтов А.Д., Передерий А.А., Охлопкова О.В., Саушкина Л.А., Шведова Е.В., Гришина Е.А., Лиознов Д.А., 20262026Желтухина А.И., Комиссаров А.Б., Фадеев А.В., Ёлшин Н.Д., Писарева М.М., Едер В.А., Коржанова М., Ксенафонтов А.Д., Передерий А.А., Охлопкова О.В., Саушкина Л.А., Шведова Е.В., Гришина Е.А., Лиознов Д.А.Zheltukhina A.I., Komissarov A.B., Fadeev A.V., Yolshin N.D., Pisareva M.M., Eder V.A., Korzhanova M., Ksenafontov A.D., Perederiy A.A., Ohlopkova O.V., Saushkina L.A., Shvedova E.V., Grishina E.A., Lioznov D.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.biopreparations.ru/jour/article/view/750ВВЕДЕНИЕ

ВВЕДЕНИЕ. Спонтанные мутации часто возникают и накапливаются в геноме РНК-содержащих вирусов, таких как вирусы кори, краснухи и паротита. Современные требования к производству вакцинных препаратов, содержащих живые вакцинные штаммы вирусов, предполагают контроль генетической стабильности производственных штаммов. Метод высокопроизводительного секвенирования позволяет выявить все мутации, возникшие в вирусной популяции, и оценить генетическую стабильность производственных штаммов.

ЦЕЛЬ

ЦЕЛЬ. Анализ нуклеотидных последовательностей геномов посевных и производственных штаммов вирусов кори (Ленинград-16), краснухи (RA-27/3) и паротита (Ленинград-3) в сравнении с референсными нуклеотидными последовательностями геномов вакцинных штаммов, депонированных ранее в базу данных GenBank.

МАТЕРИАЛЫ И МЕТОДЫ

МАТЕРИАЛЫ И МЕТОДЫ. В работе использовали посевные вирусы и производственные штаммы вакцин: коревая культуральная живая, паротитная культуральная живая и вакцина против краснухи культуральная живая. Проведено полногеномное секвенирование с использованием генетических анализаторов MGI DNBSEQ G-400 и Illumina NextSeq2000. Для обработки и анализа данных секвенирования использовали программы и инструменты FastQC v.0.12.1, FastP v.0.23.4, BWA-MEM2 v.2.2.1, VizCoV.

РЕЗУЛЬТАТЫ

РЕЗУЛЬТАТЫ. Анализ распространения однонуклеотидных полиморфизмов (SNP) внутри вирусной популяции в исследованных образцах выявил ряд минорных вариантов, способных при дальнейшем накоплении привести к изменению консенсусных последовательностей белков вакцинных штаммов. В образцах вируса кори не выявлено полиморфизмов в белке Н, при этом обнаружен несинонимичный полиморфизм в белке F, встречающийся в 30,0–40,9% вирусных геномов. В капсидном белке С и в поверхностном гликопротеине Е2 вируса краснухи выявлены полиморфизмы, встречающиеся в вирусной популяции с частотой до 30%. В поверхностных антигенах — гемагглютинин-нейраминидаза HN и белок слияния F вируса паротита обнаружены полиморфизмы, встречающиеся в геномах менее чем в 20% вирусной популяции. Мутации в геномах производственных штаммов, которые могли бы оказывать влияние на иммуногенность вакцин против кори, краснухи и паротита, не найдены.

ВЫВОДЫ

ВЫВОДЫ. Выявленный набор минорных вариантов с частотой встречаемости в популяции более 10%, которые при дальнейшем накоплении могут привести к изменению консенсусных последовательностей вакцинных штаммов, актуализирует задачу контроля стабильности вакцинных штаммов методом высокопроизводительного секвенирования.

INTRODUCTION

INTRODUCTION. Spontaneous mutations often arise and accumulate in the genome of RNA-containing viruses, for example measles, rubella and mumps. Modern requirements for the production of vaccine products containing live vaccine viruses suggest control of the genetic stability of production strains. The method of high throughput sequencing allows to determine all mutations arisen in viral population, and to assess genetic stability of production strains.

AIM

AIM. Analysis of nucleotide sequence of the genomes of seed and production strains of measles (Leningrad-16), rubella (RA27/3) and mumps (Leningrad-3) viruses, in comparison with the reference nucleotide sequences of vaccine strains genomes (submitted in the Gen-Bank database).

MATERIALS AND METHODS

MATERIALS AND METHODS. Production strains of vaccines (measles culture live, rubella culture live and mumps culture live) were used in this study. Whole genome sequencing was performed using the MGI DNBSEQ G-400 and Illumina NextSeq2000 genetic analyzers. When processing and analyzing the sequencing data, the following programs and tools were used: FastQC v.0.12.1, FastP v.0.23.4, BWA-MEM2 v.2.2.1, VizCoV tool.

RESULTS

RESULTS. Analysis of single nucleotide polymorphisms (SNPs) within the viral population in the studied samples revealed a number of minor variants that, and after further accumulation, may lead to changes in the consensus sequences of the vaccine strains proteins. For the studied measles virus samples, no SNPs were found in the main measles virus antigen H protein, however, a nonsynonymous polymorphism was detected in the F protein, occurring in 30.0–40.9% of the viral genomes. SNPs were found in the capsid protein C and in the surface glycoprotein E2 rubella virus, occurring in the viral population with a frequency of up to 30%. SNPs were found in surface antigens hemagglutinin neuraminidase HN and fusion protein F of mumps virus, occurring in the genomes of less than 20% of viral population. Mutations in the genomes of production strains that could affect the immunogenicity of measles, rubella, and mumps vaccines were not detected.

CONCLUSIONS

CONCLUSIONS. The identified set of minor variants with a frequency of occurrence in viral population of more than 10%, which, with further accumulation, can lead to changes in the consensus sequences of vaccine strains, makes the monitoring of vaccine strains stability using high-throughput sequencing relevant.

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The authors declare that there are no conflicts of interest present.