Czech J. Food Sci., 2009, 27(10):S350-S353 | DOI: 10.17221/959-CJFS

Nucleic Acid Lateral Flow Immunoassay for the Detection of Pathogenic Bacteria from Food

M. Blažková*, M. Koets, J. H. Wichers, A. van Amerongen, L. Fukal, P. Rauch
Department of Biochemistry and Microbiology, Institute of Chemical Technology in Prague, 66 28 Prague, Czech Republic

Nucleic acid lateral flow immunoassay (NALFIA) is a method combining molecular biological principle of detection with immunochemical principle of visualisation. Following isolation of DNA from the sample, a duplex PCR with two primer sets, of which one was labelled with biotin and the other with digoxigenin or fluorescein, respectively, was performed. The PCR solution and carbon particles conjugated with avidin are directly added to the nitrocellulose membrane with two test lines of immobilised antibodies specific for digoxigenin and fluorescein. The appearance of a black line indicates the presence of specific amplicon. We would like to present the NALFIA for the simultaneous detection of L. monocytogenes in particular and the genus Listeria in general, in food. Bacteria from the genus Listeria frequently contaminate a large variety of foods. Occurrence of Listeria strains in food may indicate errors in good hygienic and manufacturing practice, only L. monocytogenes is a significant human and animal pathogen responsible for the serious illness listeriosis. Conventional microbiological methods for L. monocytogenes detection are laborious and take several days to achieve a confirmed identification.

Keywords: L. monocytogenes; Nucleic acid lateral flow immunoassay; DNA

Published: June 30, 2009  Show citation

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Blažková M, Koets M, Wichers JH, van Amerongen A, Fukal L, Rauch P. Nucleic Acid Lateral Flow Immunoassay for the Detection of Pathogenic Bacteria from Food. Czech J. Food Sci. 2009;27(Special Issue 1):S350-353. doi: 10.17221/959-CJFS.
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