foodsystFood systemsПищевые системы2618-97712618-7272Федеральный научный центр пищевых систем им. В.М. Горбатова РАН10.21323/2618-9771-2025-8-3-324-334foodsyst-847Research ArticleСтатьиDevelopment of multiplex PCR for species-specific DNA determination of saury (Cololabis saira) and far eastern sardine, ivasi (Sardinops melanostictus)Разработка мультиплексной ПЦР для определения видоспецифичной ДНК сайры (Cololabis saira) и дальневосточной сардины, иваси (Sardinops melanostictus)https://orcid.org/0000-0001-8848-0632ФоминаТ. А.FominaT. A.

Фомина Татьяна Алексеевна — кандидат технических наук, старший научный сотрудник, Лаборатория молекулярной биологии и биоинформатики; Начальник отдела молекулярно-диагностических исследований испытательной референс-лаборатории

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

Tatyana A. Fomina, Candidate of Technical Sciences, Senior Research Scientist, Laboratory of Molecular Biology and Bioinformatics; Head of the Department of Molecular Diagnostic Research at the Testing Reference Laboratory

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

fomina1032@yandex.ruhttps://orcid.org/0000-0002-3445-4559ХвостовД. В.KhvostovD. V.

Хвостов Даниил Владиславович — кандидат технических наук, научный сотрудник, лаборатория «Молекулярной биологии и  биоинформатики»

109316, Москва, ул. Талалихина, 26

Daniil V.  Khvostov, Candidate of Technical Sciences, Scientific Worker, Laboratory of Molecular Biology and Dioinformatics

26, Talalikhin str., 109316, Moscow

d.hvostov@fncps.ruhttps://orcid.org/0000-0002-0038-9744МинаевМ. Ю.MinaevM. Yu.

Минаев Михаил Юрьевич — кандидат технических наук, руководитель лаборатории Молекулярной биологии и  биоинформатики

109316, г. Москва, ул. Талалихина, 26

Mikhail Yu. Minaev, Candidate of Technical Sciences, Head of Laboratory of Molecular Biology and Bioinformatics

26, Talalikhin str., 109316, Moscow

m.minaev@fncps.ruhttps://orcid.org/0000-0001-7693-3032УтьяновД. А.UtyanovD. A.

Утьянов Дмитрий Александрович — кандидат технических наук, научный сотрудник, лаборатория «Научно-методические работы, биологические и аналитические исследования»

109316, Москва, ул. Талалихина,26

Dmitry A.  Utyanov, Candidate of Technical Sciences, Scientific Worker, Laboratory of Scientific and Methodical Work, Biological and Analytical Research

26, Talalikhin str., 109316, Moscow

d.utyanov@fncps.ruФедеральный научный центр пищевых систем им. В. М. Горбатова; Национальный центр безопасности рыбной и сельскохозяйственной продукцииРоссияV. M. Gorbatov Federal Research Center for Food Systems; Nation Center for Fish and Agricultural Product SafetyRussian FederationФедеральный научный центр пищевых систем им. В. М. ГорбатоваРоссияV. M. Gorbatov Federal Research Center for Food SystemsRussian Federation20251610202583324334Copyright © Fomina T.A., Khvostov D.V., Minaev M.Y., Utyanov D.A., 20252025Фомина Т.А., Хвостов Д.В., Минаев М.Ю., Утьянов Д.А.Fomina T.A., Khvostov D.V., Minaev M.Y., Utyanov 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.fsjour.com/jour/article/view/847

The paper presents the development and validation of a multiplex real-time polymerase chain reaction (real-time PCR) method forspecies identification of canned products from saury (Cololabis saira) and Far Eastern sardine (Ivasi) (Sardinops melanostictus). The method is based on the use of the oligonucleotide system «Saury-Ivasi-VKO» and is aimed at solving the problem of adulteration of fish products, which is widespread in the Russian market. To assess the specificity of the method, a control panel of 90 fish species and 195 food fish samples were used. The method demonstrated high sensitivity, allowing detection of DNA of target species at up to 0.01% in the sample. The specificity of primers was confirmed by the absence of DNA amplification of unrelated species. The conducted experiments showed that the declared fish species were correctly identified in 100% of cases, and there were no false negative results. The results of food product testing revealed nonconformities in 34% of canned saury samples, of which 21 % were replaced by sardine and 6% by Pacific herring. The developed methodology reduces the cost of analysis due to the lack of dependence on imported reagents and can be implemented in laboratories working with GMOs and species identification. The conclusions confirm the effectiveness of the proposed approach for quality control and prevention of adulteration of fish products.

В статье представлены разработка и апробация метода мультиплексной полимеразной цепной реакции в реальном времени (ПЦР-РВ) для видовой идентификации консервированной продукции из сайры (Cololabis saira) и дальневосточно сардины (иваси) (Sardinops melanostictus). Методика основана на использовании системы олигонуклеотидов «Сайра-ИвасиВКО» и направлена на решение проблемы фальсификации рыбной продукции, которая широко распространена на российском рынке. Для оценки специфичности метода была использована контрольная панель из 90 видов рыб, а также 195 образцов пищевой рыбной продукции. Метод продемонстрировал высокую чувствительность, позволяя выявлять ДНК целевых видов при содержании до 0,01% в образце. Специфичность праймеров подтверждена отсутствием амплификации ДНК неродственных видов. Проведенные эксперименты показали, что заявленные виды рыбы корректно идентифицируются в 100 % случаев, а ложноотрицательные результаты отсутствуют. Результаты исследования пищевой продукции продемонстрировали несоответствия в 34 % образцов консервов из сайры, из которых 21% заменены сардиной, а 6% — тихоокеанской сельдью. Разработанная методика снижает затраты на анализ за счет отсутствия зависимости от импортных реагентов и может быть внедрена в лабораториях, работающих с ГМО и видовой идентификацией. Выводы подтверждают эффективность предложенного подхода для контроля качества и предотвращения фальсификации рыбной продукции.

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