Malorny Burkhard, Bunge Cornelia, Guerra Beatriz, Prietz Sandra, Helmuth Reiner
National Salmonella Reference Laboratory, Federal Institute for Risk Assessment, Diedersdorfer Weg 1, D-12277 Berlin, Germany.
Mol Cell Probes. 2007 Feb;21(1):56-65. doi: 10.1016/j.mcp.2006.08.005. Epub 2006 Sep 17.
A DNA microarray has been developed for the simultaneous characterisation and typing of Salmonella enterica subsp. enterica isolates. One-hundred and nine 35-40 mer oligonucleotides probes detect flagellar and somatic antigen encoding genes (serogroup or serotype specific), important virulence genes located within or outside the pathogenicity islands, phage-associated genes and antibiotic resistance determinants. The probes were printed on glass slides and whole genomic Cy5-labelled Salmonella DNA was hybridised to the substrate. A set of 19 different Salmonella strains and one Escherichia coli strain has been selected as positive and negative controls for each probe. The validity of the results is confirmed by gene-specific PCRs or phenotypic methods (serotyping, MIC determination for various antimicrobial agents). Of 2071 data points generated, an agreement of 97.4% has been obtained between microarray and PCR/phenotypic results. Twenty-six data points (1.3%) were classified as uncertain and, similarly, 1.3% showed a discordant result. The microarray described here is a new tool to study the epidemiology of Salmonella strains on the genotypic level and might become a powerful method in risk assessment studies.
已开发出一种DNA微阵列，用于同时鉴定和分型肠炎沙门氏菌亚种肠炎分离株。109个35 - 40聚体寡核苷酸探针可检测鞭毛和菌体抗原编码基因（血清群或血清型特异性）、位于致病岛内外的重要毒力基因、噬菌体相关基因和抗生素抗性决定簇。这些探针被印制在玻片上，全基因组Cy5标记的沙门氏菌DNA与该基质杂交。已选择一组19种不同的沙门氏菌菌株和1种大肠杆菌菌株作为每种探针的阳性和阴性对照。结果的有效性通过基因特异性PCR或表型方法（血清分型、各种抗菌剂的最低抑菌浓度测定）得到证实。在产生的2071个数据点中，微阵列与PCR/表型结果之间的一致性达到了97.4%。26个数据点（1.3%）被归类为不确定，同样，1.3%显示出不一致的结果。这里描述的微阵列是在基因型水平上研究沙门氏菌菌株流行病学的一种新工具，可能会成为风险评估研究中的一种强大方法。
