Zou Wen, Al-Khaldi Sufian F, Branham William S, Han Tao, Fuscoe James C, Han Jing, Foley Steven L, Xu Joshua, Fang Hong, Cerniglia Carl E, Nayak Rajesh
Division of Personalized Nutrition and Medicine, National Center for Toxicological Research, Food and Drug Administration, Jefferson, AR 72079, United States.
J Infect Dev Ctries. 2011 Mar 2;5(2):94-105. doi: 10.3855/jidc.1396.
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.
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.
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.
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.
对食源性病原体致病潜力进行快速、准确且低成本的分析是食品安全和生物防御领域的一项重要考量，在发展中国家尤为如此。本研究的目的是展示使用一种强大且低成本的微阵列平台来检测食品和/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。
点阵微阵列由69个选定的沙门氏菌特异性毒力基因探针（每个65bp）组成。这些探针被打印在聚-L-赖氨酸包被的载玻片上。基因组DNA用Sau3AI酶切，用Cy3染料标记，与基因探针杂交，然后用GenePix 4000B和ArrayTrack™（美国食品药品监督管理局（FDA）研究人员开发的一款免费软件）采集并分析图像。
在所检测的所有沙门氏菌菌株中，近58%的毒力相关基因都存在。一般来说，属于inv、pip、prg、sic、sip、spa或ttr家族的基因在超过90%的分离株中被检测到，而iacP、avrA、invH、rhuM、sirA、sopB、sopE或sugR基因在40%至80%的分离株中被检测到。基因变异性与沙门氏菌血清型无关。
这种杂交微阵列通过针对一组特定的沙门氏菌相关毒力基因，为在疫情调查中评估沙门氏菌的致病潜力提供了一种准确且经济高效的方法。
