Microarray analysis of microbial virulence factors
Microarray analysis of microbial virulence factors
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DOI：
10.1128/aem.67.7.3258-3263.2001
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发表时间：
2001-07-01
影响因子：
4.4
通讯作者：
Levy, DD
中科院分区：
文献类型：
--
作者：
Chizhikov, V;Rasooly, A;Levy, DD
寡核苷酸微芯片（微阵列）杂交用于大肠杆菌和其他致病性肠道细菌的菌株之间的歧视窝藏各种毒力因子。寡核苷酸微芯片是固定在玻璃表面上的基因特异性寡核苷酸探针的微型阵列。该技术与PCR扩增遗传物质相结合，是检测和同时区分食源性人类病原体的有力工具。通过多重PCR监测编码细菌抗原决定簇和细菌菌株毒力因子的六个基因（eae-1、slt-I、slt-II、fliC、rfbE和ipaH）的存在，然后将变性PCR产物与微芯片上的基因特异性寡核苷酸杂交。该检测方法能够检测15种沙门氏菌、志贺氏菌和大肠杆菌菌株中的这些毒力因子。芯片分析的结果通过放射性标记的基因特异性探针与来自细菌菌落的基因组DNA的杂交来证实。相比之下，用于微阵列分析的多重PCR产物的凝胶电泳分析由于意外和未表征的条带的存在而产生模糊的结果。我们的研究结果表明，微阵列分析的微生物毒力因子可能是非常有用的自动化鉴定和表征的细菌病原体。
Hybridization with oligonucleotide microchips (microarrays) was used for discrimination among strains of Escherichia coli and other pathogenic enteric bacteria harboring various virulence factors. Oligonucleotide microchips are miniature arrays of gene-specific oligonucleotide probes immobilized on a glass surface. The combination of this technique with the amplification of genetic material by PCR is a powerful tool for the detection of and simultaneous discrimination among food-borne human pathogens. The presence of six genes (eae-l, slt-I, slt-II,fliC, rfbE, and ipaH) encoding bacterial antigenic determinants and virulence factors of bacterial strains was monitored by multiplex PCR followed by hybridization of the denatured PCR product to the gene-specific oligonucleotides on the microchip. The assay was able to detect these virulence factors in 15 Salmonella, Shigella, and E, coli strains, The results of the chip analysis were confirmed by hybridization of radiolabeled gene-specific probes to genomic DNA from bacterial colonies. In contrast, gel electrophoretic analysis of the multiplex PCR products used for the microarray analysis produced ambiguous results due to the presence of unexpected and uncharacterized bands. Our results suggest that microarray analysis of microbial virulence factors might be very useful for automated identification and characterization of bacterial pathogens.