Abstract
Detection of pathogenic bacteria that pose a great risk to human health requires a rapid, convenient, reliable, and sensitive detection method. In this study, we developed a selective filtration method using monoclonal antibody (MAb)–magnetic nanoparticle (MNP) nanocomposites for the rapid and sensitive colorimetric detection of Salmonella typhimurium. The method contains two key steps: the immunomagnetic separation of the bacteria using MAb–MNP nanocomposites and the filtration of the nanocomposite-bound bacteria. Color signals from the nanocomposites remaining on the membrane were measured, which reflected the amount of bacteria in test samples. Immunomagnetic capture efficiencies of 8 to 90 % for various concentrations of the pathogen (2 × 104–2 × 101 cells) were obtained. After optimization of the method, 2 × 101 cells of S. typhimurium in pure culture solution was detectable as well as in artificially inoculated vegetables (100 cells/g). The method was confirmed to be highly specific to S. typhimurium without cross-reaction to other pathogenic bacteria and could be concluded within 45 min, yielding results in a shorter or similar time period as compared with recently reported antibody immobilized on magnetic-particle-based methods. This study also demonstrated direct application of MAb–MNP nanocomposites without a dissociation step of bacteria from magnetic beads in colorimetric assays in practice.
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Acknowledgments
This work was financially supported by a grants from the NLRL Program (2001-0028915) and the Converging Research Center Program (2013K000242) through the National Research Foundation of Korea (NRF) funded by the Korean Ministry of Science, ICT & Future Planning and by the Fishery Commercialization Technology Development Program, Korean Ministry of Oceans and Fisheries.
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Won-Bo Shim and Jeong-Eon Song contributed equally to this work.
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Shim, WB., Song, JE., Mun, H. et al. Rapid colorimetric detection of Salmonella typhimuriumusing a selective filtration technique combined with antibody–magnetic nanoparticle nanocomposites. Anal Bioanal Chem 406, 859–866 (2014). https://doi.org/10.1007/s00216-013-7497-6
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DOI: https://doi.org/10.1007/s00216-013-7497-6