vestichИзвестия Национальной академии наук Беларуси. Серия химических наукProceedings of the National Academy of Sciences of Belarus, Chemical Series1561-83312524-2342The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8331-2026-62-2-132-141vestich-1016Research ArticleБИООРГАНИЧЕСКАЯ ХИМИЯBIOORGANIC CHEMISTRYКонструкция и аналитические характеристики новой иммуноферментной системы для обнаружения Listeria monocytogenes в молокеDesign and analytical characteristics of a new enzyme-linked immunosorbent assay system for the detection of Listeria monocytogenes in milkКиселеваЕ. П.KiselevaE. P.

Киселева Елена Павловна – кандидат химических наук, ведущий научный сотрудник.

Ул. Купревича, 5/2, 220084, Минск

Kiseleva Elena P. – Ph. D. (Chemistry), Leading Researcher.

5/2, Kuprevich Str., 220084, Minsk

epkiseleva@iboch.byМихайлопулоК. И.MikhailopuloK. I.

Михайлопуло Константин Игоревич – старший научный сотрудник.

Ул. Купревича, 5/2, 220084, Минск

Mikhailopulo Konstantin I. – Senior Researcher.

5/2, Kuprevich Str., 220141, Minsk

k.mikhailopulo@iboch.byСвиридовО. В.SviridovO. V.

Свиридов Олег Васильевич – доктор химических наук, профессор, заведующий лабораторией.

Ул. Купревича, 5/2, 220084, Минск

Sviridov Oleg V. – Dr. Sci. (Chemistry), Professor, Head of Laboratory.

5/2, Kuprevich Str., 220084, Minsk

sviridov@iboch.byИнститут биоорганической химии Национальной академии наук БеларусиБеларусьInstitute of Bioorganic Chemistry of the National Academy of Sciences of BelarusBelarus202626052026622132141Copyright © Киселева Е.П., Михайлопуло К.И., Свиридов О.В., 20262026Киселева Е.П., Михайлопуло К.И., Свиридов О.В.Kiseleva E.P., Mikhailopulo K.I., Sviridov O.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestichem.belnauka.by/jour/article/view/1016

Разработана тест-система для обнаружения Listeria monocytogenes в культуральной среде, основанная на принципе конкурентного иммуноферментного анализа и имеющая аналитическую чувствительность 1 ∙ 104 КОЕ/мл. Тест-система отличается от известных аналогов (коммерческих и описанных в научных исследованиях) двумя конструктивными особенностями: цитозоль клеток Listeria monocytogenes (мембранно-свободная фракция) иммобилизован на твердой фазе; анализируемым биоматериалом является среда, в которой выращивались бактерии. Адаптация данной тест-системы к иммуноферментному анализу Listeria monocytogenes в молоке выявила выраженный матричный эффект, который не устранялся традиционными методами. При добавлении молока в калибровочные пробы непосредственно перед проведением иммуноферментного анализа сохранена аналитическая чувствительность, равная 1 ∙ 104 КОЕ/мл.

Detection of Listeria monocytogenes (LM) in foods is topical due to known cases of listeriosis epidemics. Immunochemical methods including ELISA are widely used for LM detection. Traditionally, commercial ELISA kits for LM detection in foods are based on sandwich technique, detect antigens of bacterial cell membrane, have analytical sensitivity ~ 105 CFU/mL, and the analyzed biomaterial is cells + medium. We elaborated competitive ELISA test for LM detection in foods with analytical sensitivity 1  104 CFU/mL. The test system differs from known analogues (commercial and described in research studies) by two design features: the membrane-free fraction of LM cells (cytosol) is immobilized on the solid |phase; the analyzed biomaterial is the medium in which the bacteria were grown. Adaptation of this test system to the ELISA detection of LM in milk revealed a pronounced matrix effect that could not be eliminated by conventional methods. We addressed this issue by adding milk to the calibration samples immediately before performing the ELISA, maintaining an analytical sensitivity of 1 · 104 CFU/mL.

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