Advertisement

Analytical and Bioanalytical Chemistry

, Volume 407, Issue 24, pp 7285–7294 | Cite as

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

  • Luis Antonio Tortajada-Genaro
  • Alejandro Rodrigo
  • Elizabeth Hevia
  • Salvador Mena
  • Regina Niñoles
  • Ángel Maquieira
Research Paper

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.

Graphical Abstract

DNA microarray performed by DVD technology for pathogen genotyping

Keywords

Genotyping Food-borne pathogens DNA assay Compact disc Microarray 

Notes

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.

References

  1. 1.
    European Food Safety Authority (2014) EFSA J 12:3547Google Scholar
  2. 2.
    Gould LH, Walsh KA, Vieira AR, Herman K, Williams IT, Hall AJ, Cole D (2013) Morb Mortal Wkly Rep (MMWR) 62:1–34Google Scholar
  3. 3.
    ISO 6579:2002, Microbiology of food and animal feeding stuffs. Horizontal method for the detection of Salmonella sppGoogle Scholar
  4. 4.
    ISO 10272–1:2006, Microbiology of food and animal feeding stuffs. Horizontal method for detection and enumeration of Campylobacter spp. Part 1: detection methodGoogle Scholar
  5. 5.
    Kim S, Frye JG, Hu J, Fedorka-Cray PJ, Gautom R, Boyle DS (2006) J Clin Microbiol 44:3608–3615CrossRefGoogle Scholar
  6. 6.
    Jasson V, Jacxsens L, Luning P, Rajkovic A, Uyttendaele M (2010) Food Microbiol 27:710–730CrossRefGoogle Scholar
  7. 7.
    Park SH, Aydin M, Khatiwara A, Dolan MC, Gilmore DF, Bouldin JL, Ahn S, Ricke SC (2014) Food Microbiol 38:250–262CrossRefGoogle Scholar
  8. 8.
    Lazcka O, Del Campo FJ, Munoz FX (2007) Biosens Bioelectron 22:1205–1217CrossRefGoogle Scholar
  9. 9.
    Velusamy V, Arshak K, Korostynska O, Oliwa K, Adley C (2010) Biotechnol Adv 28:232–254CrossRefGoogle Scholar
  10. 10.
    Wang Y, Ye Z, Ying Y (2012) Sensors (Basel) 12:3449–3471CrossRefGoogle Scholar
  11. 11.
    Mairhofer J, Roppert K, Ertl P (2009) Sensors (Basel) 9:4804–4823CrossRefGoogle Scholar
  12. 12.
    Hsieh K, Patterson AS, Ferguson BS, Plaxco KW, Soh HT (2012) Angew Chem Int Ed 51:4896–4900CrossRefGoogle Scholar
  13. 13.
    Donhauser SC, Niessner R, Seidel M (2011) Anal Chem 83:3153–3160CrossRefGoogle Scholar
  14. 14.
    Vora GJ, Meador CE, Stenger DA, Andreadis JD (2004) Appl Environ Microbiol 70:3047–3054CrossRefGoogle Scholar
  15. 15.
    Wang XW, Zhang L, Jin LQ, Jin M, Shen ZQ, An S, Chao FH, Li JW (2007) Appl Environ Microbiol 76:225–233Google Scholar
  16. 16.
    Lopez-Campos G, Martínez-Suarez JV, Aguado-Urda M, Lopez-Alonso V (2012) Microarray detection and characterization of bacterial foodborne pathogens. Springer, New YorkCrossRefGoogle Scholar
  17. 17.
    Suo B, He Y, Paoli G, Gehring A, Tu SI, Shi X (2010) Mol Cell Probes 24:77–86CrossRefGoogle Scholar
  18. 18.
    Gronlund H, Riber L, Vigre H, Lofstrom C, Folling L, Huehn S, Malorny B, Radstrom P, Rudi K, Hoorfar J (2011) Int J Food Microbiol 145:S79–S85CrossRefGoogle Scholar
  19. 19.
    Arnandis-Chover T, Morais S, Tortajada-Genaro LA, Puchades R, Maquieira A, Berganza J, Olabarria G (2012) Talanta 101:405–412CrossRefGoogle Scholar
  20. 20.
    Santiago-Felipe S, Tortajada-Genaro LA, Morais S, Puchades R, Maquieira A (2015) Food Chem 174:509–515CrossRefGoogle Scholar
  21. 21.
    Jacxsens L, Kussaga J, Luning PA, Van der Spiegel M, Devlieghere F, Uyttendaele M (2009) Int J Food Microbiol 134:113–125CrossRefGoogle Scholar
  22. 22.
    Sethi D, Gandhi RP, Kuma P, Gupta KC (2009) Biotechnol J 4:1513–1529CrossRefGoogle Scholar
  23. 23.
    Kim HJ, Park SH, Lee TH, Nahm BH, Kim YR, Kim HY (2008) Biosens Bioelectron 24:238–246CrossRefGoogle Scholar
  24. 24.
    Guo D, Liu B, Liu F, Cao B, Chen M, Hao X, Feng L, Wang L (2013) Appl Environ Microbiol 79:3392–3399CrossRefGoogle Scholar
  25. 25.
    Smith CJ, Osborn AM (2009) FEMS Microbiol Ecol 67:6–20CrossRefGoogle Scholar
  26. 26.
    Cheung PY, Kam KM (2012) Food Res Int 45:802–808CrossRefGoogle Scholar
  27. 27.
    Feng J, Wang X, Cao G, Hu S, Kuang X, Tang S, You S, Liu L (2013) Eur Food Res Technol 236:1073–1083CrossRefGoogle Scholar
  28. 28.
    Suo B, He Y, Irwin P, Gehring A (2013) Food Anal Methods 6:1477–1484CrossRefGoogle Scholar
  29. 29.
    Shin HH, Hwang BH, Seo JH, Cha HJ (2014) Appl Environ Microbiol 80:366–373CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Luis Antonio Tortajada-Genaro
    • 1
  • Alejandro Rodrigo
    • 2
  • Elizabeth Hevia
    • 1
  • Salvador Mena
    • 1
  • Regina Niñoles
    • 1
  • Ángel Maquieira
    • 1
  1. 1.Instituto Int. Reconocimiento Molecular (IDM), Departamento de QuímicaUniversitat Politècnica de ValènciaValenciaSpain
  2. 2.Asociación de Investigación de la Industria Agroalimentaria (AINIA)PaternaSpain

Personalised recommendations