foodsystFood systemsПищевые системы2618-97712618-7272Федеральный научный центр пищевых систем им. В.М. Горбатова РАН10.21323/2618-9771-2022-5-2-80-93foodsyst-154Research ArticleСтатьиIdentification of fish species using the next generation sequencing (NGS) technologyИдентификация видов рыбы с помощью технологии секвенирования нового поколения (NGS)https://orcid.org/0000-0001-8848-0632ФоминаТ. А.FominaT. A.

МоскваФомина Татьяна Алексеевна — кандидат технических наук, старший научный сотрудник, Лаборатория молекулярной биологии и биоинформатики, Федеральный научный центр пищевых систем им. В. М. Горбатова; Начальник отдела молекулярно-диагностических исследований испытательной референс-лаборатории, Национальный центр безопасности продукции водного промысла и аквакультуры109316, Москва, ул. Талалихина, 26; 129626, Москва, Графский переулок, д.14–1

Tatyana A. Fomina, Candidate of Technical Sciences, Senior Research Scientist, Laboratory of Molecular Biology and Bioinformatics, V. M. Gorbatov Federal Research Center for Food Systems; Head of the Department of Molecular Diagnostic Research at the Testing Reference Laboratory, National Centre for Safety of Aquatic Fisheries Products and Aquaculture

26, Talalikhina str., 109316, Moscow; 14–1, Grafskiy per., 129626, Moscow

https://orcid.org/0000-0001-5581-7534КулешоваМ. Г.KuleshovaM. G.

Кулешова Мария Геннадьевна — аспирант, Лаборатория молекулярной биологии и биоинформатики, Федеральный научный центр пищевых систем им. В. М. Горбатова; Молекулярный биолог, Отдел молекулярно-диагностических исследований испытательной референс-лаборатории, Национальный центр безопасности продукции водного промысла и аквакультуры

109316, Москва, ул. Талалихина, 26; 129626, Москва, Графский переулок, д.14–1

Mariya G.  Kuleshova, Graduate Student, Laboratory of Molecular Biology and Bioinformatics, V. M. Gorbatov Federal Research Center for Food Systems; Molecular Biologist, Department of Molecular Diagnostic Research at the Testing Reference Laboratory, National Centre for Safety of Aquatic Fisheries Products and Aquaculture

26, Talalikhina str., 109316, Moscow; 14–1, Grafskiy per., 129626, Moscow

mariakulg@mail.ruhttps://orcid.org/0000-0002-0038-9744МинаевМ. Ю.MinaevM. Yu.

Минаев Михаил Юрьевич — кандидат технических наук, руководитель лаборатории Молекулярной биологии и  биоинформатики, Федеральный научный центр пищевых систем им. В. М. Горбатова

109316, Москва, ул. Талалихина, 26

Mikhail Yu. Minaev, Candidate of Technical Sciences, Head of Laboratory of Molecular Biology and Bioinformatics, V. M. Gorbatov Federal Research

Center for Food Systems 26, Talalikhina str., 109316, Moscow

m.minaev@fncps.ruhttps://orcid.org/0000-0001-8748-9117КоноровЕ. А.KonorovE. A.

Коноров Евгений Андреевич — кандидат биологических наук, старший научный сотрудник, Лаборатория молекулярной биологии и биоинформатики, Федеральный научный центр пищевых систем им. В. М. Горбатова

109316, Москва, ул. Талалихина, 26

Evgeniy А. Konorov, Candidate of Technical Sciences, Senior Research Scientist, Laboratory of Molecular Biology and Bioinformatics, V. M. Gorbatov Federal Research Center for Food Systems

26, Talalikhina str., 109316, Moscow

casqy@yandex.ruФедеральный научный центр пищевых систем им. В. М. Горбатова; Национальный центр безопасности продукции водного промысла и аквакультурыРоссияV. M. Gorbatov Federal Research Center for Food Systems; National Centre for Safety of Aquatic Fisheries Products and AquacultureRussian FederationФедеральный научный центр пищевых систем им. В. М. ГорбатоваРоссияV. M. Gorbatov Federal Research Center for Food SystemsRussian Federation202208072022528093Copyright © Fomina T.A., Kuleshova M.G., Minaev M.Y., Konorov E.A., 20222022Фомина Т.А., Кулешова М.Г., Минаев М.Ю., Коноров Е.А.Fomina T.A., Kuleshova M.G., Minaev M.Y., Konorov E.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.fsjour.com/jour/article/view/154

The laws relating to fish and fishery product labeling that require indication of the information about fish species exist in many world countries. These rules are conditioned by a significant growth in the number of the economic fraud cases in the field of production and trade of fishery products. The widespread ways of fraud are replacement and mislabeling of a product as confirmed by many studies. Analysis of scientific works shows that mislabeling in fishery product manufacture occurs in 30–70% of cases in different countries. The existing legislation about food traceability is insufficient for their prevention, which suggests a necessity of taking strict control measures ensuring effective species identification of fish and fishery products. At present, various laboratory tests are used for their species identification. They are based, mainly, on analysis of unique DNA profiles found in different species. In this work, we present the method for detection of fish species using next generation sequencing (NGS). NGS is an advanced technology in the field of quality control of fishery products, especially for fish species identification in multicomponent products, which contain DNA fragments of other species besides the target DNA. NGS was carried out on the platform Ion Torrent Ion GeneStudio S5 System. Twenty samples were analyzed: 17 commercial samples and three prepared experimental samples consisted of the mixture of two and more species. The universal primers, which were able to amplify the fragment 16S rRNA of the commercial fish species, were selected and prepared. In general, DNA of 11 families, 15 genera and 16 species was identified in the course of the analysis. The obtained result of NGS of 17 commercial samples confirmed the results of identification by other molecular diagnostic methods. Mislabeling was revealed in four samples. In three samples, all fish species present in the composition were identified. Possible reasons for fish replacement were assessed.

Законы о маркировке рыбы и рыбной продукции, требующие указывать информацию о видовой принадлежности рыбы, есть во многих странах мира. Данные правила обусловлены высоким ростом количества случаев экономического мошенничества в области производства и оборота рыбных продуктов. Распространенными способами мошенничества являются подмена и  неправильная маркировка продукта, что подтверждается многочисленными исследованиями. Анализ научных работ показал, что применение неправильной маркировки при производстве рыбных продуктов в разных странах встречается в 30–70% случаев. Для их предотвращения имеющегося законодательства о прослеживаемости пищевых продуктов недостаточно, что указывает на необходимость принятия строгих мер контроля, обеспечивающих эффективную видовую идентификацию рыбы и рыбопродуктов. В настоящее время для идентификации их видовой принадлежности используют различные лабораторные тесты, главным образом основанные на анализе уникальных профилей ДНК, которые обнаружены у различных видов. В данной работе представлен метод определения видовой принадлежности рыб с использованием секвенирования нового поколения (NGS). Технология секвенирования NGS является передовой в области контроля качества рыбной продукции, особенно для идентификации видов рыб в многокомпонентной продукции, в которой помимо целевой ДНК присутствуют и фрагменты ДНК других видов. Секвенирование NGS проводили на платформе Ion Torrent Ion GeneStudio S5 System, были проанализированы двадцать образцов, 17 коммерческих образцов и 3 приготовленных опытных образца, состоящих их смеси двух и более видов. Были подобраны и подготовлены универсальные праймеры, способные амплифицировать фрагмент 16S rRNA промысловых видов рыб. В целом в ходе анализа была идентифицирована ДНК 11 семейств, 15 родов и 16 видов. Полученный результат секвенирования NGS17 коммерческих образцов подтверждал результаты идентификации другими молекулярно-диагностическими методами, была выявлена неправильная маркировка в 4 образцах. В 3 опытных образцах были идентифицированы все виды рыб, входящих в их состав. Была произведена оценка возможных причин замены рыбы.

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