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Applied and Environmental Microbiology, November 2009, p. 6647-6654, Vol. 75, No. 21
0099-2240/09/$08.00+0     doi:10.1128/AEM.00854-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Multiplexed Bead-Based Mesofluidic System for Detection of Food-Borne Pathogenic Bacteria {triangledown}

Sheng-Quan Jin, Bin-Cheng Yin, and Bang-Ce Ye*

Lab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, China

Received 15 April 2009/ Accepted 23 August 2009

In the present study, a simple and rapid multiplexed bead-based mesofluidic system (BMS) was developed for simultaneous detection of food-borne pathogenic bacteria, including Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella, Enterobacter sakazakii, Shigella, Escherichia coli O157:H7, and Campylobacter jejuni. This system is based on utilization of isothiocyanate-modified microbeads that are 250 µm in diameter, which were immobilized with specific amino-modified oligonucleotide probes and placed in polydimethylsiloxane microchannels. PCR products from the pathogens studied were pumped into microchannels to hybridize with the oligonucleotide-modified beads, and hybridization signals were detected using a conventional microarray scanner. The short sequences of nucleic acids (21 bases) and PCR products characteristic of bacterial pathogens could be detected at concentrations of 1 pM and 10 nM, respectively. The detection procedure could be performed in less than 30 min with high sensitivity and specificity. The assay was simple and fast, and the limits of quantification were in the range from 500 to 6,000 CFU/ml for the bacterial species studied. The feasibility of identification of food-borne bacteria was investigated with samples contaminated with bacteria, including milk, egg, and meat samples. The results demonstrated that the BMS method can be used for effective detection of multiple pathogens in different foodstuffs.

* Corresponding author. Mailing address: Lab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, China. Phone and fax: 86-21-64252094. E-mail: bcye{at}ecust.edu.cn

{triangledown} Published ahead of print on 28 August 2009.

Applied and Environmental Microbiology, November 2009, p. 6647-6654, Vol. 75, No. 21
0099-2240/09/$08.00+0     doi:10.1128/AEM.00854-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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