A Subtractively Optimized DNA Microarray Using Non-sequenced Genomic Probes for the Detection of Food-Borne Pathogens |
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Authors: | Jin Yong Lee Byoung Chan Kim Kwan Jong Chang Joo-Myung Ahn Jee-Hoon Ryu Hyo-Ihl Chang Man Bock Gu |
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Institution: | (1) School of Life Sciences and Biotechnology, Korea University, Anam-dong, Seongbuk-gu, Seoul, 136-701, Republic of Korea;(2) Environment Division, Korea Institute of Science and Technology, Hawolgok-dong, Seongbuk-gu, Seoul, 136-791, Republic of Korea; |
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Abstract: | In this study, we present the successful detection of food-borne pathogens using randomly selected non-sequenced genomic DNA
probes-based DNA microarray chips. Three food-borne pathogens, Staphylococcus aureus, Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium), and Bacillus cereus, were subjected for the preparation of the DNA microarray probes. Initially, about 50 DNA probes selected randomly from non-sequenced
genomic DNA of each pathogen were prepared by using a set of restriction enzyme pairs. The proto-type of DNA microarray chip
for detecting three different pathogens simultaneously was fabricated by using those DNA probes prepared for each pathogen.
This proto-type DNA microarray has been tested with three target pathogens and additional seven bacteria, and successfully
verified with a few cross-hybridized probes. After this primary verification of the DNA microarray hybridization, this proto-type
DNA microarray chip was redesigned and successfully optimized by eliminating a few cross-hybridized probes. The specificity
of this redesigned DNA microarray chip to each pathogen was confirmed without any serious cross-hybridizations, and its multiplexing
capability in its pathogen detection was found to be possible. This randomly selected non-sequenced genomic DNA probes-based
DNA microarray was successfully proved to be the high-throughput simultaneous detection chip for the detection of food-borne
pathogens, without knowing the exact sequence information of the target bacteria. This could be the first fabrication of DNA
microarray chip for the simultaneous detection of different kinds of food-borne pathogens. |
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