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Microarray on digital versatile disc for identification and genotyping of Salmonella and Campylobacter in meat products

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Abstract

Highly portable, cost-effective, and rapid-response devices are required for the subtyping of the most frequent food-borne bacteria; thereby the sample rejection strategies and hygienization techniques along the food chain can be tailor-designed. Here, a novel biosensor is presented for the generic detection of Salmonella and Campylobacter and the discrimination between their most prevalent serovars (Salmonella Enteritidis, Salmonella Typhimurium) and species (Campylobacter jejuni, Campylobacter coli), respectively. The method is based on DNA microarray developed on a standard digital versatile disc (DVD) as support for a hybridization assay and a DVD driver as scanner. This approach was found to be highly sensitive (detection limit down to 0.2 pg of genomic DNA), reproducible (relative standard deviation 4–19 %), and high working capacity (20 samples per disc). The inclusivity and exclusivity assays indicated that designed oligonucleotides (primers and probes) were able to discriminate targeted pathogens from other Salmonella serovars, Campylobacter species, or common food-borne pathogens potentially present in the indigenous microflora. One hundred isolates from meat samples, collected in a poultry factory, were analyzed by the DVD microarraying and fluorescent real-time PCR. An excellent correlation was observed for both generic and specific detection (relative sensitivity 93–99 % and relative specificity 93–100 %). Therefore, the developed assay has been shown to be a reliable tool to be used in routine food safety analysis, especially in settings with limited infrastructure due to the excellent efficiency-cost ratio of compact disc technology.

DNA microarray performed by DVD technology for pathogen genotyping

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Acknowledgments

This work has been supported by the Spanish Ministry of Economy and Competitiveness projects:INNPACTO (Ref. IPT-2011-1011-31000) and FEDER-CTQ2013-45875-R). Contribution of grant PROMETEO II/2014/040 (Generalitat Valenciana) was highly acknowledged.

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Authors and Affiliations

  1. Instituto Int. Reconocimiento Molecular (IDM), Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022, Valencia, Spain

    Luis Antonio Tortajada-Genaro, Elizabeth Hevia, Salvador Mena, Regina Niñoles & Ángel Maquieira

  2. Asociación de Investigación de la Industria Agroalimentaria (AINIA), Benjamín Franklin 5-11, 46980, Paterna, Valencia, Spain

    Alejandro Rodrigo

Authors
  1. Luis Antonio Tortajada-Genaro
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  2. Alejandro Rodrigo
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  3. Elizabeth Hevia
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  4. Salvador Mena
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  5. Regina Niñoles
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  6. Ángel Maquieira
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Corresponding author

Correspondence to Ángel Maquieira.

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Tortajada-Genaro, L.A., Rodrigo, A., Hevia, E. et al. Microarray on digital versatile disc for identification and genotyping of Salmonella and Campylobacter in meat products. Anal Bioanal Chem 407, 7285–7294 (2015). https://doi.org/10.1007/s00216-015-8890-0

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  • DOI: https://doi.org/10.1007/s00216-015-8890-0

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