Microarray detection of food-borne pathogens using specific probes prepared by comparative genomics.
In order to design a method for the accurate detection and identification of food-borne pathogens, we used comparative genomics to select 70-mer oligonucleotide probes specific for 11 major food-borne pathogens (10 overlapping probes per pathogen) for use in microarray analysis. We analyzed the hybridization pattern of this constructed microarray with the Cy3-labeled genomic DNA of various food-borne pathogens and other bacteria. Our microarray showed a highly specific hybridization pattern with the genomic DNA of each food-borne pathogen; little unexpected cross-hybridization was observed. Microarray data were analyzed and clustered using the GenePix Pro 6.0 and GeneSpring GX 7.3.1 programs. The analyzed dendrogram revealed the discriminating power of constructed microarray. Each food-borne pathogen clustered according to its hybridization specificity and non-pathogenic species were discriminated from pathogenic species. Our method can be applied to the rapid and accurate detection and identification of food-borne pathogens in the food industry. In addition, this study demonstrates that genome sequence comparison and DNA microarray analysis have a powerful application in epidemiologic and taxonomic studies, as well as in the food safety and biodefense fields.
Kim
Hyun-Joong
HJ
Institute of Life Sciences & Resources and Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.
Park
Si-Hong
SH
Lee
Tae-Ho
TH
Nahm
Baek-Hie
BH
Kim
Young-Rok
YR
Kim
Hae-Yeong
HY
D002874:Chromosome Mapping / Q000379:methods*; D015342:DNA Probes / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D005504:Food Analysis / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*
D015342:DNA Probes; D004269:DNA, Bacterial