S H Ohk1, O K Koo, T Sen, C M Yamamoto, A K Bhunia. 1. Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47907-2009, 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 invasive 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.
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 invasive 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.
Authors: John G Bruno; Taylor Phillips; Tiffany Montez; Adrian Garcia; Jeffrey C Sivils; Michael W Mayo; Alex Greis Journal: J Fluoresc Date: 2014-12-17 Impact factor: 2.217