Detection of Representative Enteropathogenic Bacteria, Vibrio spp., Pathogenic Escherichia coli, Salmonella spp., Shigella spp., and Yersinia enterocolitica, Using a Virulence Factor Gene-Based Oligonucleotide Microarray§

Dong-Hun Kim; Bok-Kwon Lee; Yong-Dae Kim; Sung-Keun Rhee; Young-Chang Kim

doi:10.1007/s12275-010-0119-5

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Detection of Representative Enteropathogenic Bacteria, Vibrio spp., Pathogenic Escherichia coli, Salmonella spp., Shigella spp., and Yersinia enterocolitica, Using a Virulence Factor Gene-Based Oligonucleotide Microarray§: Dong-Hun Kim ¹, Bok-Kwon Lee ², Yong-Dae Kim ³, Sung-Keun Rhee ^1†, Young-Chang Kim ^1†; Journal of Microbiology 2010;48(5):682-688.
DOI: https://doi.org/10.1007/s12275-010-0119-5
Published online: November 3, 2010

¹Department of Microbiology, Chungbuk National University, Cheongju 361-763, Republic of Korea, ²Division of Enteric Bacterial Infections, Center for Infectious Diseases, Korea National Institute of Health, Seoul 112-701, Republic of Korea, ³Department of Preventive Medicine, Chungbuk National University, Cheongju 361-763, Republic of Korea

Corresponding author: Young-Chang Kim , Tel: +82-43-261-2302,

Received: 31 March 2010 • Accepted: 19 May 2010

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Abstract

Rapid identification of enteropathogenic bacteria in stool samples is critical for clinical diagnosis and antimicrobial therapy. In this study, we describe the development of an approach that couples multiplex PCR with hybridization to a DNA microarray, to allow the simultaneous detection of the 10 pathogens. The microarray was synthesized with 20-mer oligonucleotide probes that were designed to be specific for virulencefactor genes of each strain. The detection limit for genomic DNA from a single strain was approximately 10 fg. In the presence of heterogeneous non-target DNA, the detection sensitivity of the array decreased to approximately 100 fg. We did not observe any non-specific hybridization. In addition, we successfully used this oligonucleotide-based DNA microarray to identify the causative agents in clinical stool samples from patients with food-borne enteritis.

Keywords: multiplex PCR;microarray;oligonucleotide probes;enteropathogenic bacteria

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Detection of Representative Enteropathogenic Bacteria, Vibrio spp., Pathogenic Escherichia coli, Salmonella spp., Shigella spp., and Yersinia enterocolitica, Using a Virulence Factor Gene-Based Oligonucleotide Microarray§

J. Microbiol. 2010;48(5):682-688. Published online November 3, 2010

DOI: https://doi.org/10.1007/s12275-010-0119-5

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