Design and analytical characteristics of a new enzyme-linked immunosorbent assay system for the detection of Listeria monocytogenes in milk
https://doi.org/10.29235/1561-8331-2026-62-2-132-141
Abstract
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.
About the Authors
E. P. Kiseleva
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus
Belarus
Belarus
Kiseleva Elena P. – Ph. D. (Chemistry), Leading Researcher.
5/2, Kuprevich Str., 220084, Minsk
K. I. Mikhailopulo
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus
Belarus
Belarus
Mikhailopulo Konstantin I. – Senior Researcher.
5/2, Kuprevich Str., 220141, Minsk
O. V. Sviridov
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus
Belarus
Belarus
Sviridov Oleg V. – Dr. Sci. (Chemistry), Professor, Head of Laboratory.
5/2, Kuprevich Str., 220084, Minsk
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