Electrokinetically-controlled RNA-DNA hybridization assay for foodborne pathogens
- 160 Downloads
We have developed a microfluidic chip for use in an RNA-DNA hybridization assay for foodborne pathogens. Automatic sequential reagent dispensing and washing was realized with a programmable DC voltage sequencer. Signal detection was achieved with a miniaturized optical detection module. Salmonella and Listeria monocytogenes bacteria in different concentrations were quantitatively determined by this RNA-DNA hybridization assay in the microfluidic chip. The detection limit for the Salmonella and Listeria monocytogenes bacteria is 103 to 104 CFU mL−1. The method excels by a significant reduction in the consumption of sample and reagent, and a short assay time. This automatic-operating microfluidic RNA-DNA hybridization assay is promising for on-site pathogen detection.
KeywordsRNA-DNA hybridization assay Electrokinetics Microfluidics Salmonella Listeria monocytogenes
The authors would like to express appreciation to the financial support of the Canada Research Chairs program and the Natural Sciences and Engineering Research Council (NSERC) of Canada through a research grant to Dr. Dongqing Li. Thanks also go to Dr. Shu Chen and Dr. Honghe Cao (Laboratory Services Division, University of Guelph) for providing the reagents and bacterial samples.
- 5.Artault S, Blind JL, Delaval J, Dureuil Y, Gaillard N (2001) Detecting Listeria monocytogenes in food. Int Food Hyg 12:23Google Scholar
- 7.Valdivieso-Garcia A, Desruisseau A, Riche E, Fukuda S, Tatsumi H (2003) Evaluation of a 24- hour bioluminescent enzyme immunoassay for the rapid detection of Salmonella in chicken carcass rinses. J Food Prot 66:1996–2004Google Scholar
- 9.Magliulo M, Simoni P, Guardigli M, Michelini E, Luciani M, Lelli R, Roda A (2007) A rapid multiplexed chemiluminescent immunoassay for the detection of Escherichia coli O157: H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes pathogen bacteria. J Agric Food Chem 55:4933–4939CrossRefGoogle Scholar
- 11.Lei IF, Roffey P, Blanchard C, Gu K (2008) Development of a multiplex PCR method for the detection of six common foodborne pathogens. J Food Drug Anal 16:37–43Google Scholar
- 12.Kim JS, Lee GG, Park JS, Jung YH, Kwak HS, Kim SB, Nam YS, Kwon ST (2007) A novel multiplex PCR assay for rapid and simultaneous detection of five pathogenic bacteria: Escherichia coli O157: H7, Salmonella, Staphylococcus aureus, Listeria monocytogenes, and Vibrio parahaemolyticus. J Food Protect 70:1656–1662Google Scholar