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Antibody–aptamer functionalized fibre‐optic biosensor for specific detection of Listeria monocytogenes from food
Authors:SH Ohk  OK Koo  T Sen  CM Yamamoto  AK Bhunia
Institution:1. Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA;2. Hitachi Chemical Research Center, Inc., Irvine, CA, USA
Abstract: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. 103 CFU ml?1. This sensor also successfully detected L. monocytogenes cells from artificially contaminated (initial inoculation of 102 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.
Keywords:aptamer  detection  fibre‐optic sensor  food  Listeria monocytogenes
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