microbeПроблемы особо опасных инфекцийProblems of Particularly Dangerous Infections0370-10692658-719XRussian Research Anti-Plague Institute “Microbe”10.21055/0370-1069-2021-2-94-99microbe-1500Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESРазработка иммуноферментных тест-систем и иммунохроматографических наборов реагентов, предназначенных для выявления стафилококковых энтеротоксинов типов А и ВDevelopment of Elisa Test-Systems and Immune-Chromatographic Reagent Panel Designed for the Detection of Staphylococcal Enterotoxins of A and B Typeshttps://orcid.org/0000-0003-3133-1921ЕремкинА. В.EremkinA. V.

Kirov

https://orcid.org/0000-0002-2881-4730ИпатовС. С.IpatovS. S.

Kirov

https://orcid.org/0000-0002-5429-6295КуклинаГ. В.KuklinaG. V.

Kirov

https://orcid.org/0000-0001-8277-3573ПеченкинД. В.PechenkinD. V.

Kirov

23527@mil.ruhttps://orcid.org/0000-0003-0277-2226КытмановА. А.KytmanovA. A.

Kirov

https://orcid.org/0000-0001-8368-9648ТихвинскаяО. В.TikhvinskayaO. V.

Kirov

https://orcid.org/0000-0003-2705-8774КузнецовС. Л.KuznetsovS. L.

Kirov

https://orcid.org/0000-0001-5164-3645ГоршковА. С.GorshkovA. S.

Kirov

https://orcid.org/0000-0002-9201-9803ХапаевН. Г.KhapaevN. G.

Kirov

Филиал ФГБУ «48 Центральный научно-исследовательский институт» Министерства обороны Российской ФедерацииРоссияAffiliated Branch of the Federal State Budgetary Institution “48th Central Research Institute” of the Ministry of Defense of the Russian FederationRussian Federation202121072021029499Copyright © Еремкин А.В., Ипатов С.С., Куклина Г.В., Печенкин Д.В., Кытманов А.А., Тихвинская О.В., Кузнецов С.Л., Горшков А.С., Хапаев Н.Г., 20212021Еремкин А.В., Ипатов С.С., Куклина Г.В., Печенкин Д.В., Кытманов А.А., Тихвинская О.В., Кузнецов С.Л., Горшков А.С., Хапаев Н.Г.Eremkin A.V., Ipatov S.S., Kuklina G.V., Pechenkin D.V., Kytmanov A.A., Tikhvinskaya O.V., Kuznetsov S.L., Gorshkov A.S., Khapaev N.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.microbe.ru/jour/article/view/1500Цель

Цель. Создание экспериментальных образцов иммуноферментных тест-систем и иммунохроматографических наборов реагентов для выявления стафилококковых энтеротоксинов типов А и В.

Материалы и методы

Материалы и методы. В работе использованы гибридомы, продуцирующие моноклональные антитела к стафилококковым энтеротоксинам типов А и В, полученные в филиале ФГБУ «48 ЦНИИ» Минобороны России (г. Киров); мыши линии BALB/c; стафилококковые энтеротоксины типов А и В. Клетки гибридом культивировали в культуральных флаконах и в перитонеальной полости мышей линии BALB/с. Моноклональные антитела выделяли из асцитических жидкостей путем осаждения насыщенным раствором сульфата аммония с последующей очисткой ионообменной хроматографией. Полученные препараты моноклональных антител использовали для конструирования иммуноферментных тестсистем и иммунохроматографических наборов реагентов для выявления стафилококковых энтеротоксинов типов А и В. Специфические компоненты иммуноферментных тест-систем подвергали сублимационному высушиванию в защитной среде.

Результаты и обсуждение

Результаты и обсуждение. Сконструированы иммуноферментные тест-системы и иммунохроматографические наборы реагентов, позволяющие выявлять стафилококковые энтеротоксины типов А и В в концентрациях 0,5 нг/мл и более, в том числе в пробах продуктов питания. 

Objective of the study was the development of experimental ELISA tests and lateral flow immunoassays for detection of staphylococcal enterotoxins, A and B types.

Materials and methods

Materials and methods. Hybridomas, producing monoclonal antibodies against staphylococcal enterotoxins A and B from State Collection of the Affiliated Branch of the “48th Central Research Institute” of the Ministry of Defense of the Russian Federation, BALB/c mice and staphylococcal enterotoxins A and B were used in the research. Hybridoma cells were incubated in culture flasks and in the peritoneal cavity of BALB/c mice. Monoclonal antibodies were isolated from ascitic fluids through precipitation with saturated ammonium sulfate subsequently purified using ion-exchange chromatography. Obtained preparations of monoclonal antibodies were used for the construction of ELISA tests and immune-chromatographic reagent panels for the detection of staphylococcal enterotoxins A and B. Specific components of ELISA tests were lyophilized in protective media.

Results and discussion

Results and discussion. ELISA tests and lateral flow immunoassays which allow for detecting staphylococcal enterotoxins A and B at concentrations of 0.5 ng/ml and higher, including in food samples, have been constructed. 

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