van Hoek Angela H A M, Scholtens Ingrid M J, Cloeckaert Axel, Aarts Henk J M
RIKILT-Institute of Food Safety, Bornsesteeg 45, NL-6708PD Wageningen, The Netherlands.
J Microbiol Methods. 2005 Jul;62(1):13-23. doi: 10.1016/j.mimet.2005.01.004.
In this study the feasibility of 50- and 60-mer oligonucleotides in microarray analysis for the detection and identification of antibiotic resistance genes in various Salmonella strains was assessed. The specificity of the designed oligonucleotides was evaluated, furthermore the optimal spotting concentration was determined. The oligonucleotide microarray was used to screen two sets of Salmonella strains for the presence of several antibiotic resistance genes. Set 1 consisted of strains with variant Salmonella Genomic Island 1 (SGI1) multidrug resistance (MDR) regions of which the antibiotic resistance profiles and genotypes were known. The second set contained strains of which initially only phenotypic data were available. The microarray results of the first set of Salmonella strains perfectly matched with the phenotypic and genotypic information. The microarray data of the second set were almost completely in concordance with the available phenotypic data. It was concluded that the microarray technique in combination with random primed genomic labeling and 50- or 60-mer oligonucleotides is a powerful tool for the detection of antibiotic resistance genes in bacteria.
在本研究中，评估了50聚体和60聚体寡核苷酸在微阵列分析中用于检测和鉴定各种沙门氏菌菌株中抗生素抗性基因的可行性。评估了所设计寡核苷酸的特异性，此外还确定了最佳点样浓度。寡核苷酸微阵列用于筛选两组沙门氏菌菌株中几种抗生素抗性基因的存在情况。第一组由具有不同沙门氏菌基因组岛1（SGI1）多药耐药（MDR）区域的菌株组成，其抗生素抗性谱和基因型是已知的。第二组包含最初仅具有表型数据的菌株。第一组沙门氏菌菌株的微阵列结果与表型和基因型信息完全匹配。第二组的微阵列数据几乎与可用的表型数据完全一致。得出的结论是，微阵列技术结合随机引物基因组标记和50聚体或60聚体寡核苷酸是检测细菌中抗生素抗性基因的有力工具。
