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

• Published in: The journal of microbiology Vol. 48; no. 5; pp. 682 - 688
• Main Authors: Kim, D.H., Chungbuk National University, Cheongju, Republic of Korea, Lee, B.K., Korea National Institute of Health, Seoul, Republic of Korea, Kim, Y.D., Chungbuk National University, Cheongju, Republic of Korea, Rhee, S.K., Chungbuk National University, Cheongju, Republic of Korea, Kim, Y.C., Chungbuk National University, Cheongju, Republic of Korea
• Format: Journal Article
• Language: English
• Published: Heidelberg The Microbiological Society of Korea 01.10.2010; Springer Nature B.V; 한국미생물학회
• Subjects: Bacteria; Bacterial infections; Bacteriological Techniques - methods; Biomedical and Life Sciences; DNA, Bacterial - genetics; DNA, Bacterial - isolation & purification; E coli; Enterobacteriaceae - genetics; Enterobacteriaceae - pathogenicity; Enterobacteriaceae Infections - diagnosis; Enterobacteriaceae Infections - microbiology; enteropathogenic bacteria; Escherichia coli; Feces - microbiology; Foodborne Diseases - diagnosis; Foodborne Diseases - microbiology; Genes; Genomics; Health care; Humans; Hybridization; Infectious diseases; Life Sciences; microarray; Microbiology; multiplex PCR; Oligonucleotide Array Sequence Analysis - methods; oligonucleotide probes; Pathogens; PCR; Polymerase Chain Reaction - methods; Preventive medicine; Salmonella; Sensitivity and Specificity; Shigella; Vibrio; Virulence; Virulence Factors - genetics; Virulence Factors - isolation & purification; Yersinia enterocolitica; 생물학; United States > US; South Korea; multiplex PCR; microarray; enteropathogenic bacteria; oligonucleotide probes
• ISSN: 1225-8873; 1976-3794
• DOI: 10.1007/s12275-010-0119-5

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 1.0 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.