A Subtractively Optimized DNA Microarray Using Non-sequenced Genomic Probes for the Detection of Food-Borne Pathogens

被引：1

作者：

Lee, Jin Yong ^{[1
]}

Kim, Byoung Chan ^{[2
]}

Chang, Kwan Jong ^{[1
]}

Ahn, Joo-Myung ^{[1
]}

Ryu, Jee-Hoon ^{[1
]}

Chang, Hyo-Ihl ^{[1
]}

Gu, Man Bock ^{[1
]}

机构：

[1] Korea Univ, Sch Life Sci & Biotechnol, Seoul 136701, South Korea

[2] Korea Inst Sci & Technol, Environm Div, Seoul 136791, South Korea

来源：

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | 2011年 / 164卷 / 02期

关键词：

Food-borne pathogen; DNA microarray chip; Non-sequenced genomic DNA probes; Cross-hybridization; Multiplexed detection; GRADIENT GEL-ELECTROPHORESIS; FOODBORNE PATHOGENS; ESCHERICHIA-COLI; IDENTIFICATION; BACTERIA; PCR; HYBRIDIZATION; MILK; DGGE;

D O I：

10.1007/s12010-010-9126-6

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：183 / 193

页数：11

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