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      Development of gold nanoparticle-aptamer-based LSPR sensing chips for the rapid detection of Salmonella typhimurium in pork meat

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          Abstract

          A non-labeled, portable plasmonic biosensor-based device was developed to enable the ultra-sensitive and selective detection of Salmonella typhimurium in pork meat samples. Specifically, a plasmonic sensor, using the self-assembly of gold nanoparticles (AuNPs) to achieve a regulated diameter of 20 nm for the AuNP monolayers, was used to conduct high-density deposition on a transparent substrate, which produced longitudinal wavelength extinction shifts via a localized surface plasmon resonance (LSPR) signal. The developed aptamers conjugated to the LSPR sensing chips revealed an ultra-sensitive upper limit of detection (LOD) of approximately 10 4 cfu/mL for S. typhimurium in pure culture under the optimal assay conditions, with a total analysis time of 30–35 min. When the LSPR sensing chips were applied on artificially contaminated pork meat samples, S. typhimurium in the spiked pork meat samples was also detected at an LOD of 1.0 × 10 4 cfu/mL. The developed method could detect S. typhimurium in spiked pork meat samples without a pre-enrichment step. Additionally, the LSPR sensing chips developed against S. typhimurium were not susceptible to any effect of the food matrix or background contaminant microflora. These findings confirmed that the developed gold nanoparticle-aptamer-based LSPR sensing chips could facilitate sensitive detection of S. typhimurium in food samples.

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          Most cited references38

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          Kinetically controlled seeded growth synthesis of citrate-stabilized gold nanoparticles of up to 200 nm: size focusing versus Ostwald ripening.

          Monodisperse citrate-stabilized gold nanoparticles with a uniform quasi-spherical shape of up to ∼200 nm and a narrow size distribution were synthesized following a kinetically controlled seeded growth strategy via the reduction of HAuCl(4) by sodium citrate. The inhibition of any secondary nucleation during homogeneous growth was controlled by adjusting the reaction conditions: temperature, gold precursor to seed particle concentration, and pH. This method presents improved results regarding the traditional Frens method in several aspects: (i) it produces particles of higher monodispersity; (ii) it allows better control of the gold nanoparticle size and size distribution; and (iii) it leads to higher concentrations. Gold nanoparticles synthesized following this method can be further functionalized with a wide variety of molecules, hence this method appears to be a promising candidate for application in the fields of biomedicine, photonics, and electronics, among others. © 2011 American Chemical Society
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            Label-free impedimetric biosensor for Salmonella Typhimurium detection based on poly [pyrrole-co-3-carboxyl-pyrrole] copolymer supported aptamer.

            The Gram-negative bacterium, Salmonella Typhimurium (S. Typhimurium) is a food borne pathogen responsible for numerous hospitalisations and deaths all over the world. Conventional detection methods for pathogens are time consuming and labour-intensive. Hence, there is considerable interest in faster and simpler detection methods. Polypyrrole-based polymers, due to their intrinsic chemical and electrical properties, have been demonstrated to be valuable candidates for the fabrication of chemo/biosensors and functional surfaces. Similarly aptamers have been shown to be good alternatives to antibodies in the development of affinity biosensors. In this study, we report on the combination of poly [pyrrole-co-3-carboxyl-pyrrole] copolymer and aptamer for the development of a label-less electrochemical biosensor suitable for the detection of S. Typhimurium. Impedimetric measurements were facilitated by the effect of the aptamer/target interaction on the intrinsic conjugation of the poly [pyrrole-co-3-carboxyl-pyrrole] copolymer and subsequently on its electrical properties. The aptasensor detected S. Typhimurium in the concentration range 10(2)-10(8) CFU mL(-1) with high selectivity over other model pathogens and with a limit of quantification (LOQ) of 100 CFU mL(-1) and a limit of detection (LOD) of 3 CFU mL(-1). The suitability of the aptasensor for real sample detection was demonstrated via recovery studies performed in spiked apple juice samples. We envisage this to be a viable approach for the inexpensive and rapid detection of pathogens in food, and possibly in other environmental samples.
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              An aptamer-based electrochemical biosensor for the detection of Salmonella.

              Salmonella is one of the most common causes of food-associated disease. An electrochemical biosensor was developed for Salmonella detection using a Salmonella-specific recognition aptamer. The biosensor was based on a glassy carbon electrode modified with graphene oxide and gold nanoparticles. Then, the aptamer ssDNA sequence could be linked to the electrode. Each assembly step was accompanied by changes to the electrochemical parameters. After incubation of the modified electrode with Salmonella, the electrochemical properties between the electrode and the electrolyte changed accordingly. The electrochemical impedance spectrum was measured to quantify the Salmonella. The results revealed that, when more Salmonella were added to the reaction system, the current between the electrode and electrolyte decreased; in other words, the impendence gradually increased. A detection limit as low as 3 cfu/mL was obtained. This novel method is specific and fast, and it has the potential for real sample detection.
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                Author and article information

                Contributors
                smyoo@kaist.ac.kr
                yunsuk.huh@inha.ac.kr
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                31 August 2017
                31 August 2017
                2017
                : 7
                : 10130
                Affiliations
                [1 ]ISNI 0000 0001 2364 8385, GRID grid.202119.9, Department of Chemical Engineering, Inha University, 100 Inha-ro, Nam-gu, ; Incheon, 22212 Republic of Korea
                [2 ]ISNI 0000 0001 0671 5021, GRID grid.255168.d, Department of Energy and Materials Engineering, Dongguk University-Seoul, 30 Pildong-ro 1-gil, ; Seoul, 04620 Republic of Korea
                [3 ]ISNI 0000 0001 2296 8192, GRID grid.29869.3c, Reliability Assessment Center for Chemical Materials, Korea Research Institute of Chemical Technology (KRICT), 141 Gajeong-ro, Yuseong-gu, ; Daejeon, 34114 Republic of Korea
                [4 ]ISNI 0000 0001 2292 0500, GRID grid.37172.30, Department of Chemical and Biomolecular Engineering (BK21 plus program), KAIST, ; Daejeon, 34141 Republic of Korea
                [5 ]ISNI 0000 0001 0789 9563, GRID grid.254224.7, School of Integrative Engineering, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, ; Seoul, 06974 Republic of Korea
                Author information
                http://orcid.org/0000-0003-0599-3091
                Article
                10188
                10.1038/s41598-017-10188-2
                5579046
                28860462
                b58c5866-192e-45a2-9438-2cdea95aa6a1
                © The Author(s) 2017

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 17 May 2017
                : 3 August 2017
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