nbsprotВестник войск РХБ защитыJournal of NBC Protection Corps2587-57283034-279127 Научный центр10.35825/2587-5728-2021-5-4-366-375vkxfldnbsprot-93Research ArticleБиологическая безопасность и защита от биологических угрозBiological Security and Protection against Biological ThreatsПЦР-амплифицированный иммуноанализ (иммуно-ПЦР): принцип метода, варианты исполнения, возможности и перспективы использования для выявления патогенных биологических агентовPCR-amplified Immunoassay (Immuno-PCR): Principle of the Method, Variants of Execution, Possibilities and Prospects of Use for the Detection of Pathogenic Biological AgentsГоршковА. С.GorshkovA. S.

Горшков Антон Сергеевич. Научный сотрудник научно-исследовательского отдела, канд. мед. наук. 

610000, г. Киров, Октябрьский проспект, д. 119

Anton Sergeevich Gorshkov. Researcher of the Scientific and Research Department. Candidate of Medical Sciences.

Oktyabrsky Avenue 119, Kirov 610000

23527@mil.ruПеченкинД. В.PechenkinD. V.

Печенкин Денис Валериевич. Начальник научно-исследовательского отдела, канд. мед. наук.

610000, г. Киров, Октябрьский проспект, д. 119

Denis Valerievich Pechenkin. Chief of the Scientific and Research Department. Candidate of Medical Sciences.

Oktyabrsky Avenue 119, Kirov 610000

23527@mil.ruКузнецовскийА. В.KuznetsovskiyA. V.

Кузнецовский Андрей Владимирович. Начальник отдела планирования НИР – заместитель начальника филиала по НИР, канд. биол. наук.

610000, г. Киров, Октябрьский проспект, д. 119

Andrey Vladimirovich Kuznetsovskiy. Chief of the Department of Planning of Science and Research – Deputy Chief of the Branch. Candidate of Biological Sciences.

Oktyabrsky Avenue 119, Kirov 610000

23527@mil.ruБалакинВ. А.BalakinV. A.

Балакин Василий Альбертович. Инженер военного представительства.

610025, г. Киров, ул. Мельничная, д. 31

Vasiliy Albertovich Balakin. Military Engineer.

Mel'nichnaya street, 31, Kirov 610025

23527@mil.ruФилиал федерального государственного бюджетного учреждения «48 Центральный научно-исследовательский институт» Министерства обороны Российской ФедерацииРоссияBranch Office of the Federal State Budgetary Establishment «48 Central Scientific Researche Institute» of the Ministry of Defence of the Russian FederationRussian Federation2724 военное представительство Министерства обороны Российской ФедерацииРоссия724 Military Representation of the Ministry of Defence of the Russian FederationRussian Federation20210806202354366375Copyright © Горшков А.С., Печенкин Д.В., Кузнецовский А.В., Балакин В.А., 20232023Горшков А.С., Печенкин Д.В., Кузнецовский А.В., Балакин В.А.Gorshkov A.S., Pechenkin D.V., Kuznetsovskiy A.V., Balakin V.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.nbsprot.ru/jour/article/view/93

«Золотым стандартом» выявления биологических патогенов на сегодняшний день являются иммуноферментный анализ и полимеразная цепная реакция. Объединить оба метода в единую платформу, сохранить их преимущества, добиться высокой чувствительности анализа позволяет метод амплифицированного иммуноанализа – иммуно-ПЦР. Цель работы – рассмотреть возможности и перспективы использования ПЦР-амплифицированного иммуноанализа для выявления патогенных биологических агентов. Иммуно-ПЦР позволяет обнаруживать различные антигенные детерминанты ненуклеиновой природы в ПЦР за счет амплификации ДНК-метки, конъюгированной со специфическим антителом. Регистрация результата при этом также возможна в режиме реального времени по аналогии с тест-системами «real time» ПЦР. Основными методическими вопросами в технологии иммуно-ПЦР являются: выбор носителя комплексов биомолекул, выбор метода конъюгации антител детекции и репортерной нуклеиновой кислоты, оптимизация способов амплификации сигнальной ДНК и учета результатов, разработка способов снижения фоновых показателей. Мы считаем целесообразным проведение научно-исследовательских и опытно-конструкторских работ по разработке и созданию диагностических наборов реагентов на основе иммуно-ПЦР. Применительно к задаче обнаружения малых и следовых количеств антигенов патогенных биологических агентов, наиболее вероятной диагностической «нишей» метода иммуно-ПЦР будет выявление токсинов микробного и немикробного происхождения, минимальная клинически значимая доза для которых меньше чувствительности соответствующих иммунохимических тест-систем. С учетом перспектив развития метода, в будущем возможна разработка таких тест-систем для выявления аналитов-гаптенов, например некоторых токсикантов небиологического происхождения.

Enzyme immunoassay and polymerase chain reaction have become the «gold standard» for the detection of biological pathogens. The method of amplified immunoassay – immuno-PCR allows to combine both methods into a single platform to preserve their advantages and to achieve high sensitivity of the analysis. The purpose of this work is to consider the possibilities and prospects of using PCR-amplified immunoassay for the detection of pathogenic biological agents. Immuno-PCR makes it possible to detect various non-nucleic antigenic determinants in PCR by amplifying a DNA tag conjugated with a specific antibody. The registration of the results is also possible in real time as in the real-time PCR test systems. The main methodological issues in the immuno-PCR technology are: the choice of a carrier of biomolecule complexes, the choice of a method for conjugation of detection antibodies and a reporter nucleic acid, optimization of methods for amplifying signal DNA and accounting for results, and development of methods for reducing background indicators. We consider it necessary to carry out research and development work on the development and the creation of diagnostic kits based on immuno-PCR. With regard to the task of detecting small and trace amounts of antigens of pathogenic biological agents, the most likely diagnostic «niche» of the immuno-PCR method will be the detection of toxins of microbial and non-microbial origin, the minimum clinically significant dose for which is less than the sensitivity of the corresponding immunochemical test systems. Taking into account the prospects for the development of the method, in future it is possible to develop such test systems for the detection of hapten analytes, for example, some toxicants of non-biological origin.

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