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Antibody–aptamer functionalized fibre‐optic biosensor for specific detection of Listeria monocytogenes from food
Author(s) -
Ohk S.H.,
Koo O.K.,
Sen T.,
Yamamoto C.M.,
Bhunia A.K.
Publication year - 2010
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2010.04709.x
Subject(s) - listeria monocytogenes , aptamer , biosensor , polyclonal antibodies , chemistry , alexa fluor , microbiology and biotechnology , bacteria , chromatography , antibody , biology , fluorescence , biochemistry , genetics , physics , quantum mechanics , immunology
Aim: To develop antibody–aptamer functionalized fibre‐optic biosensor for specific detection of Listeria monocytogenes from food products. Methods and Results: Aptamer, a single‐stranded oligonucleotide ligand that displays affinity for the target molecule, was used in the assay to provide sensor specificity. Aptamer‐A8, specific for internalin A, an invasin protein of L. monocytogenes , was used in the fibre‐optic sensor together with antibody in a sandwich format for detection of L. monocytogenes from food. Biotinylated polyclonal anti‐ Listeria antibody, P66, was immobilized on streptavidin‐coated optical waveguide surface for capturing bacteria, and Alexa Fluor 647‐conjugated A8 was used as a reporter. The biosensor was able to selectively detect pathogenic Listeria in pure culture and in mixture with other bacteria at a concentration of approx. 10 3 CFU ml −1 . This sensor also successfully detected L. monocytogenes cells from artificially contaminated (initial inoculation of 10 2 CFU 25 g −1 ) ready‐to‐eat meat products such as sliced beef, chicken and turkey after 18 h of enrichment. Conclusion: Based on the data presented in this study, the antibody–aptamer functionalized fibre‐optic biosensor could be used as a detection tool for sensitive and specific detection of L. monocytogenes from foods. Significance and Impact of the Study: The study demonstrates feasibility and novel application of aptamer on fibre‐optic biosensor platform for the sensitive detection of L. monocytogenes from food products.