In situ DNA amplification with magnetic primers for the electrochemical detection of food pathogens

A. Lermo, S. Campoy, J. Barbé, S. Hernández, S. Alegret, M. I. Pividori

Research output: Contribution to journalArticleResearchpeer-review

110 Citations (Scopus)

Abstract

A sensitive and selective genomagnetic assay for the electrochemical detection of food pathogens based on in situ DNA amplification with magnetic primers has been designed. The performance of the genomagnetic assay was firstly demonstrated for a DNA synthetic target by its double-hybridization with both a digoxigenin probe and a biotinylated capture probe, and further binding to streptavidin-modified magnetic beads. The DNA sandwiched target bound on the magnetic beads is then separated by using a magneto electrode based on graphite-epoxy composite. The electrochemical detection is finally achieved by an enzyme marker, anti-digoxigenin horseradish peroxidase (HRP). The novel strategy was used for the rapid and sensitive detection of polymerase chain reaction (PCR) amplified samples. Promising resultants were also achieved for the DNA amplification directly performed on magnetic beads by using a novel magnetic primer, i.e., the up PCR primer bound to magnetic beads. Moreover, the magneto DNA biosensing assay was able to detect changes at single nucleotide polymorphism (SNP) level, when stringent hybridization conditions were used. The reliability of the assay was tested for Salmonella spp., the most important pathogen affecting food safety. © 2006 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)2010-2017
JournalBiosensors and Bioelectronics
Volume22
Issue number9-10
DOIs
Publication statusPublished - 15 Apr 2007

Keywords

  • Electrochemical DNA biosensor
  • HACCP
  • Magnetic primer
  • Real-time PCR
  • Salmonella spp
  • SNPs

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