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"10.29235/1561-8331-2025-61-2-141-153vestich-954Research ArticleБИООРГАНИЧЕСКАЯ ХИМИЯBIOORGANIC CHEMISTRYСистемы конкурентого иммунофлуориметрического анализа бактерий Salmonella enterica, включающие конъюгаты антител с хелатом европияSystems of competitive immunofluorometric assay of Salmonella enterica bacteria, comprising conjugates of antibodies with europium chelateСерченяТ. С.SerchenyaT. S.

Серченя Татьяна Сергеевна – кандидат химимических наук, доцент, ведущий научный сотрудник.

ул. Академика В.Ф. Купревича, 5/2, 220084, Минск

Tatyana S. Serchenya – Ph. D. (Chemistry), Associate Professor, Leading Researcher, Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus.

5/2, Academician V. F. Kuprevich Str., 220084, Minsk

serchenya@iboch.byКосмачА. А.KasmachA. A.

Космач Анастасия Александровна – младший научный сотрудник.

ул. Академика В.Ф. Купревича, 5/2, 220084, Минск

Anastasia A. Kosmach – Junior Researcher, Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus.

5/2, Academician V. F. Kuprevich Str., 220084, Minsk

kosmach_aa@iboch.byЛапинаВ. С.LapinaV. S.

Лапина Виктория Сергеевна – научный сотрудник.

ул. Академика В.Ф. Купревича, 5/2, 220084, Минск

Victoryia S. Lapina – Researcher, Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus.

5/2, Academician V. F. Kuprevich Str., 220084, Minsk

lapina@iboch.byБакаеваТ. Н.BakayevaT. N.

Бакаева Татьяна Николаевна – научный сотрудник.

ул. Филимонова, 23, 220114, Минск

Tatsiana N. Bakayeva – Researcher, Research Institute of Hygiene, Toxicology, Epidemiology, Virology and Microbiology.

23, Filimonov Str., 220114, Minsk

tanya.solo@mail.ruСвиридовО. В.SviridovO. V.

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

ул. Академика В.Ф. Купревича, 5/2, 220084, Минск

Oleg V. Sviridov – D. Sc. (Chemistry), Professor, Head of the Laboratory, Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus.

5/2, Academician V.F. Kuprevich Str., 220084

sviridov@iboch.byИнститут биоорганической химии Национальной академии наук БеларусиInstitute of Bioorganic Chemistry of the National National Academy of Sciences of BelarusНаучно-исследовательский институт гигиены, токсикологии, эпидемиологии, вирусологии и микробиологииScientific Research Institute of Hygiene, Toxicology, Epidemiology, Virology and Microbiology202529052025612141153Copyright © Серченя Т.С., Космач А.А., Лапина В.С., Бакаева Т.Н., Свиридов О.В., 20252025Серченя Т.С., Космач А.А., Лапина В.С., Бакаева Т.Н., Свиридов О.В.Serchenya T.S., Kasmach A.A., Lapina V.S., Bakayeva T.N., 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/954

Разработаны и исследованы биоаналитические системы, специфичные к бактериям Salmonella enterica и основанные на иммунохимическом связывании антигенов липополисахарида (ЛПС) этих патогенных микроорганизмов моно- и поликлональными антителами, конъюгированными с комплексонатом европия. Количественное определение клеток осуществлялось в системах иммуноанализа с детекцией на основе времяразрешенной флуоресценции (иммуноанализ с времяразрешенной флуоресценцией Eu3+, лантанидный иммунофлуоресцентный анализ, ЛИФМА, DELFIA). В микропланшетной системе ЛИФМА с меченными поликлональными антителами в растворе и белковым конъюгатом ЛПС на твердой фазе достигнуты следующие значения основных аналитических параметров: диапазон измеряемых концентраций клеток – от 104 до 107 КОЕ/мл, чувствительность IC50 – 3 · 105 КОЕ/мл, предел обнаружения (IC10) – 1,5 · 104 КОЕ/мл, коэффициент вариации результатов измерений – от 4,5 до 8,0 %. Широкая специфичность данной биоаналитической системы позволяла обнаруживать Salmonella enterica различных серотипов. Показана возможность тестирования образцов культуральной среды и молока без предварительного разведения. Степень открытия проб, содержащих S. enterica, составила 88–110 %. Представленная разработка может служить основой практического набора реагентов для контроля присутствия бактерий Salmonella enterica в пищевой продукции.

Bioanalytical systems specific to Salmonella enterica bacteria have been developed and studied. The systems are based on the immunochemical binding of lipopolysaccharide (LPS) antigens of these pathogenic microorganisms to mono- and polyclonal antibodies conjugated with a europium chelate. The quantitative determination of the cells was carried out in immunoassay systems by measuring the Eu3+ time-resolved fluorescence (dissociation-enhanced lanthanide fluorescence immunoassay, DELFIA) systems. In the DELFIA microplate system, comprising labeled polyclonal antibodies in solution and a LPS-protein conjugate on the solid-phase, the following analytical parameters were achieved: cell concentration measurement range – from 104 to 107 CFU/ml, sensitivity (IC50) – 3 · 105 CFU/ml, the limit of detection (IC10) – 1.5 · 104 CFU/ml, and the coefficient of variation – from 4,5 to 8.0 %. The broad specificity of this system enabled to detect Salmonella enterica of various serotypes. The possibility of testing samples of culture medium and milk without prior dilution was demonstrated. The recovery rate of samples containing Salmonella enterica was found to be 88–110 %. The presented development can serve as the basis for a practical kit of reagents to monitor Salmonella enterica bacteria in food products.

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