A subtractively optimized DNA microarray using non-sequenced genomic probes for the detection of food-borne pathogens.
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.
School of Life Sciences and Biotechnology, Korea University, Anam-dong, Seongbuk-gu, Seoul 136-701, Republic of Korea.
Jin Yong
JY
Lee
Byoung Chan
BC
Kim
Kwan Jong
KJ
Chang
Joo-Myung
JM
Ahn
Jee-Hoon
JH
Ryu
Hyo-Ihl
HI
Chang
Man Bock
MB
Gu
D001409:Bacillus cereus / Q000235:genetics; D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification*; D015342:DNA Probes / Q000235:genetics*; D005516:Food Microbiology / Q000379:methods*; D016680:Genome, Bacterial / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012486:Salmonella typhimurium / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D013211:Staphylococcus aureus / Q000235:genetics
D015342:DNA Probes