Microfluidic chip for direct detection of E. coli O157:H7 in ground beef via anti-digoxigenin immuno-PCR assay

Assem Abolmaaty, Hong Chen, Mohammad Faghri

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Microfluidic chip was successfully fabricated from polydimethylsiloxane (PDMS) and used for direct detection of E.coli O157:H7 in ground beef via anti-digoxigenin immuno-PCR. The PDMS channels were functionalized with 100 μg/ml streptavidin at room temperature for 6 min. A recovery of 60% ± 3.4 was obtained when bacterial cells were extracted from seeded ground beef using differential centrifugation. ProMag™ bind-IT™ microspheres were effectively used to capture 66% ± 4.2 of bacterial cells from ground beef extract. Biotinylated PCR amplicons of E. coli O157:H7 intimin gene were detected using anti-digoxigenin labeled with FITC. This chip allowed sensitive and rapid detection of a 5 cells/g ground beef while agarose gel electrophoreses failed to detect 50 cells/g ground beef. These microchannels offer pathways to eliminate the time-consuming and messy slab gel process and generate much more reproducible and high quality data.

Original languageEnglish (US)
Pages (from-to)591-598
Number of pages8
JournalWorld Applied Sciences Journal
Volume14
Issue number4
StatePublished - Dec 1 2011

All Science Journal Classification (ASJC) codes

  • General

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