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Design of a core–shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples

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Abstract

We report an immuno-magnetic separation system developed by the immobilization of pathogen-specific antibodies on the core–shell magnetic beads. The magnetic beads were grafted with glycidylmethacrylate (GMA) using surface-initiated atom transfer radical polymerization (SI-ATRP). For immuno-magnetic separation (IMS) of target bacterial cells from others, antibodies for Escherichia coli and Salmonella enterica serovar Typhimurium cells were immobilized on the magnetic beads via glutaraldehyde coupling reaction. Our IMS system successfully separated Salmonella cells when the concentrations of target (i.e., Salmonella) and interfering (i.e., E. coli) cells were at the same level. Polymerase chain reaction (PCR) assays amplifying the rfb/rfbE region of the E. coli genome and a 647-bp fragment of the invA region of Salmonella were performed as the specific selection to accurately confirm the presence of E. coli and Salmonella, respectively. IMS and multiplex PCR methods can be used for specific and quantitative detection of pathogens from food samples. Thus, this study developed a reliable and direct system for rapid detection of Salmonella and E. coli in food samples. In addition, IMS method could be easily adapted to detect other pathogens by selecting the pertinent antibody.

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Authors and Affiliations

  1. School of Medicine, Istanbul Kemerburgaz University, 34217, Istanbul, Turkey

    V. Cengiz Ozalp

  2. Biochemical Processing and Biomaterial Research Laboratory, Gazi University, 06500, Teknikokullar, Ankara, Turkey

    Gulay Bayramoglu & M. Yakup Arica

  3. Department of Chemistry, Faculty of Sciences, Gazi University, 06500, Teknikokullar, Ankara, Turkey

    Gulay Bayramoglu

  4. Department of Biology, Middle East Technical University, 06800, Ankara, Turkey

    H. Avni Oktem

Authors
  1. V. Cengiz Ozalp
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  2. Gulay Bayramoglu
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  3. M. Yakup Arica
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  4. H. Avni Oktem
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Correspondence to Gulay Bayramoglu.

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Ozalp, V.C., Bayramoglu, G., Arica, M.Y. et al. Design of a core–shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples. Appl Microbiol Biotechnol 97, 9541–9551 (2013). https://doi.org/10.1007/s00253-013-5231-0

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  • DOI: https://doi.org/10.1007/s00253-013-5231-0

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