Microarray analysis of virulence gene profiles in Salmonella serovars from food/food animal environment.
Microarray analysis of virulence gene profiles in Salmonella serovars from food/food animal environment.
复制标题
DOI：
10.3855/jidc.1396
复制
发表时间：
2011-03
影响因子：
1.9
通讯作者：
W. Zou;S. Al-Khaldi;W. Branham;T. Han;J. Fuscoe;Jing Han;S. Foley;Joshua Xu;H. Fang;C. Cer
中科院分区：
文献类型：
--
作者：
W. Zou;S. Al-Khaldi;W. Branham;T. Han;J. Fuscoe;Jing Han;S. Foley;Joshua Xu;H. Fang;C. Cer
简介快速、准确和廉价地分析食源性病原体的致病潜力是食品安全和生物防御方面的一个重要考虑因素，特别是在发展中国家。这项研究的目的是证明使用一个强大和廉价的微阵列平台来分析食品和/或食用动物环境中沙门氏菌的毒力基因谱，然后使用ArrayTrack™进行数据分析。方法斑点阵列由69个沙门氏菌特异性毒力基因探针(每个65bp)组成。这些探针被印制在涂有多聚L赖氨酸的玻片上。用Sau3AI消化基因组DNA，用Cy3染料标记，与基因探针杂交，GenePix 4000B和ArrayTrack™自由软件采集和分析图像。结果近58%的沙门氏菌毒力相关基因存在于所有沙门氏菌菌株中。90%以上的菌株检测到inv、pip、prg、sic、sip、spa或Ttr家族的基因，40%~80%的菌株检测到iacp、avra、invH、Rhum、siA、sopB、sope或SugR基因。基因变异性与沙门氏菌血清型无关。结论该杂交芯片通过选择一组沙门氏菌相关毒力基因，为在暴发调查中评估沙门氏菌的致病潜力提供了一种准确且经济有效的方法。
INTRODUCTION Rapid, accurate and inexpensive analysis of the disease-causing potential of foodborne pathogens is an important consideration in food safety and biodefense, particularly in developing countries. The objective of this study is to demonstrate the use of a robust and inexpensive microarray platform to assay the virulence gene profiles in Salmonella from food and/or the food animal environment, and then use ArrayTrack™ for data analysis. METHODOLOGY The spotted array consisted of 69 selected Salmonella-specific virulence gene probes (65bp each). These probes were printed on poly-L-lysine-coated slides. Genomic DNA was digested with Sau3AI, labeled with Cy3 dye, hybridized to the gene probes, and the images were captured and analyzed by GenePix 4000B and ArrayTrack™, a free software developed by Food and Drug Administration (FDA) researchers. RESULTS Nearly 58% of the virulence-associated genes tested were present in all Salmonella strains tested. In general, genes belonging to inv, pip, prg, sic, sip, spa or ttr families were detected in more than 90% of the isolates, while the iacP, avrA, invH, rhuM, sirA, sopB, sopE or sugR genes were detected in 40 to 80% of the isolates. The gene variability was independent of the Salmonella serotype. CONCLUSIONS This hybridization array presents an accurate and cost-effective method for evaluating the disease-causing potential of Salmonella in outbreak investigations by targeting a selective set of Salmonella-associated virulence genes.