Multiplex Solid-Phase PCR for Rapid Detection and Identification of Salmonella spp. at Sub-species

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    Abstract

    This study presents a solid-phase PCR (SP-PCR) for rapid detection, identification, and sub-typing of various Salmonella species, the major food-borne cause of salmonellosis. The target DNA is firstly amplified with PCR primers (one primer is labeled with fluorophores) in the liquid phase. Simultaneously on the solid phase, the amplified PCR amplicons interact with the nested DNA probes immobilized on the solid substrate as an array. If the immobilized probes match the sequence of the DNA templates they are extended by the polymerase and serve as template for the second strand elongation primed by the liquid phase primer thus generating new templates for the SP-PCR. After the reaction, PCR products labeled with fluorophores remain attached to the substrate and can be visualized directly by fluorescence readout devices. Using this method, S. enteritidis, S. typhimurium and S. dublin can be detected at the same time. The method offers several advantages over conventional multiplex PCR: less competition between different primer pairs thus increasing multiplexing capability, only single wavelength optical readout needed for the multiplexing detection, and less time-consuming owing to reduction of the post-PCR gel electrophoresis. The method will be useful for development of point-of-care devices for rapid detection and identification of Salmonella spp. A solid-phase PCR for rapid detection and identification of S. enteritidis, S. typhimurium and S. dublin is developed. The method offers advantages such as better multiplexing capability, only single wavelength optical readout needed, and less time-consuming.
    Original languageEnglish
    Publication date2011
    Publication statusPublished - 2011
    Event5. annual Lab-on-a-Chip European Congress - Hamburg, Germany
    Duration: 1 Jan 2011 → …
    Conference number: 5

    Conference

    Conference5. annual Lab-on-a-Chip European Congress
    Number5
    CityHamburg, Germany
    Period01/01/2011 → …

    Cite this

    Cao, C., Høgberg, J., Wolff, A., & Bang, D. D. (2011). Multiplex Solid-Phase PCR for Rapid Detection and Identification of Salmonella spp. at Sub-species. Abstract from 5. annual Lab-on-a-Chip European Congress, Hamburg, Germany, .
    @conference{58cc5093151b44768d736c6ba063cf0f,
    title = "Multiplex Solid-Phase PCR for Rapid Detection and Identification of Salmonella spp. at Sub-species",
    abstract = "This study presents a solid-phase PCR (SP-PCR) for rapid detection, identification, and sub-typing of various Salmonella species, the major food-borne cause of salmonellosis. The target DNA is firstly amplified with PCR primers (one primer is labeled with fluorophores) in the liquid phase. Simultaneously on the solid phase, the amplified PCR amplicons interact with the nested DNA probes immobilized on the solid substrate as an array. If the immobilized probes match the sequence of the DNA templates they are extended by the polymerase and serve as template for the second strand elongation primed by the liquid phase primer thus generating new templates for the SP-PCR. After the reaction, PCR products labeled with fluorophores remain attached to the substrate and can be visualized directly by fluorescence readout devices. Using this method, S. enteritidis, S. typhimurium and S. dublin can be detected at the same time. The method offers several advantages over conventional multiplex PCR: less competition between different primer pairs thus increasing multiplexing capability, only single wavelength optical readout needed for the multiplexing detection, and less time-consuming owing to reduction of the post-PCR gel electrophoresis. The method will be useful for development of point-of-care devices for rapid detection and identification of Salmonella spp. A solid-phase PCR for rapid detection and identification of S. enteritidis, S. typhimurium and S. dublin is developed. The method offers advantages such as better multiplexing capability, only single wavelength optical readout needed, and less time-consuming.",
    author = "Cuong Cao and Jonas H{\o}gberg and Anders Wolff and Bang, {Dang Duong}",
    year = "2011",
    language = "English",
    note = "5. annual Lab-on-a-Chip European Congress ; Conference date: 01-01-2011",

    }

    Cao, C, Høgberg, J, Wolff, A & Bang, DD 2011, 'Multiplex Solid-Phase PCR for Rapid Detection and Identification of Salmonella spp. at Sub-species', 5. annual Lab-on-a-Chip European Congress, Hamburg, Germany, 01/01/2011.

    Multiplex Solid-Phase PCR for Rapid Detection and Identification of Salmonella spp. at Sub-species. / Cao, Cuong; Høgberg, Jonas; Wolff, Anders; Bang, Dang Duong.

    2011. Abstract from 5. annual Lab-on-a-Chip European Congress, Hamburg, Germany, .

    Research output: Contribution to conferenceConference abstract for conferenceResearchpeer-review

    TY - ABST

    T1 - Multiplex Solid-Phase PCR for Rapid Detection and Identification of Salmonella spp. at Sub-species

    AU - Cao, Cuong

    AU - Høgberg, Jonas

    AU - Wolff, Anders

    AU - Bang, Dang Duong

    PY - 2011

    Y1 - 2011

    N2 - This study presents a solid-phase PCR (SP-PCR) for rapid detection, identification, and sub-typing of various Salmonella species, the major food-borne cause of salmonellosis. The target DNA is firstly amplified with PCR primers (one primer is labeled with fluorophores) in the liquid phase. Simultaneously on the solid phase, the amplified PCR amplicons interact with the nested DNA probes immobilized on the solid substrate as an array. If the immobilized probes match the sequence of the DNA templates they are extended by the polymerase and serve as template for the second strand elongation primed by the liquid phase primer thus generating new templates for the SP-PCR. After the reaction, PCR products labeled with fluorophores remain attached to the substrate and can be visualized directly by fluorescence readout devices. Using this method, S. enteritidis, S. typhimurium and S. dublin can be detected at the same time. The method offers several advantages over conventional multiplex PCR: less competition between different primer pairs thus increasing multiplexing capability, only single wavelength optical readout needed for the multiplexing detection, and less time-consuming owing to reduction of the post-PCR gel electrophoresis. The method will be useful for development of point-of-care devices for rapid detection and identification of Salmonella spp. A solid-phase PCR for rapid detection and identification of S. enteritidis, S. typhimurium and S. dublin is developed. The method offers advantages such as better multiplexing capability, only single wavelength optical readout needed, and less time-consuming.

    AB - This study presents a solid-phase PCR (SP-PCR) for rapid detection, identification, and sub-typing of various Salmonella species, the major food-borne cause of salmonellosis. The target DNA is firstly amplified with PCR primers (one primer is labeled with fluorophores) in the liquid phase. Simultaneously on the solid phase, the amplified PCR amplicons interact with the nested DNA probes immobilized on the solid substrate as an array. If the immobilized probes match the sequence of the DNA templates they are extended by the polymerase and serve as template for the second strand elongation primed by the liquid phase primer thus generating new templates for the SP-PCR. After the reaction, PCR products labeled with fluorophores remain attached to the substrate and can be visualized directly by fluorescence readout devices. Using this method, S. enteritidis, S. typhimurium and S. dublin can be detected at the same time. The method offers several advantages over conventional multiplex PCR: less competition between different primer pairs thus increasing multiplexing capability, only single wavelength optical readout needed for the multiplexing detection, and less time-consuming owing to reduction of the post-PCR gel electrophoresis. The method will be useful for development of point-of-care devices for rapid detection and identification of Salmonella spp. A solid-phase PCR for rapid detection and identification of S. enteritidis, S. typhimurium and S. dublin is developed. The method offers advantages such as better multiplexing capability, only single wavelength optical readout needed, and less time-consuming.

    M3 - Conference abstract for conference

    ER -

    Cao C, Høgberg J, Wolff A, Bang DD. Multiplex Solid-Phase PCR for Rapid Detection and Identification of Salmonella spp. at Sub-species. 2011. Abstract from 5. annual Lab-on-a-Chip European Congress, Hamburg, Germany, .