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利用比较基因组学制备的特异性探针进行食源性病原体的微阵列检测。

Microarray detection of food-borne pathogens using specific probes prepared by comparative genomics.

作者信息

Kim Hyun-Joong, Park Si-Hong, Lee Tae-Ho, Nahm Baek-Hie, Kim Young-Rok, Kim Hae-Yeong

机构信息

Institute of Life Sciences & Resources and Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.

出版信息

Biosens Bioelectron. 2008 Oct 15;24(2):238-46. doi: 10.1016/j.bios.2008.03.019. Epub 2008 Mar 29.

Abstract

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.

摘要

为了设计一种准确检测和鉴定食源性病原体的方法,我们利用比较基因组学选择了针对11种主要食源性病原体的70聚体寡核苷酸探针(每种病原体10个重叠探针)用于微阵列分析。我们分析了这种构建的微阵列与各种食源性病原体及其他细菌的Cy3标记基因组DNA的杂交模式。我们的微阵列与每种食源性病原体的基因组DNA呈现出高度特异性的杂交模式;几乎未观察到意外的交叉杂交。使用GenePix Pro 6.0和GeneSpring GX 7.3.1程序对微阵列数据进行分析和聚类。分析得到的树形图显示了构建的微阵列的鉴别能力。每种食源性病原体根据其杂交特异性聚类,非致病物种与致病物种得以区分。我们的方法可应用于食品工业中食源性病原体的快速准确检测和鉴定。此外,本研究表明基因组序列比较和DNA微阵列分析在流行病学和分类学研究以及食品安全和生物防御领域具有强大的应用价值。

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