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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.
MedLine Citation:
PMID:  21046348     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
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 virulence-factor 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.
Authors:
Dong-Hun Kim; Bok-Kwon Lee; Yong-Dae Kim; Sung-Keun Rhee; Young-Chang Kim
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-11-03
Journal Detail:
Title:  Journal of microbiology (Seoul, Korea)     Volume:  48     ISSN:  1976-3794     ISO Abbreviation:  J. Microbiol.     Publication Date:  2010 Oct 
Date Detail:
Created Date:  2010-11-03     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9703165     Medline TA:  J Microbiol     Country:  Korea (South)    
Other Details:
Languages:  eng     Pagination:  682-8     Citation Subset:  IM    
Affiliation:
Department of Microbiology, Chungbuk National University, Cheongju 361-763, Republic of Korea.
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