Research Center in Infectiology of Pork (CRIP), Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
Zoonoses Public Health. 2010 Nov;57 Suppl 1:94-9. doi: 10.1111/j.1863-2378.2010.01358.x.
As diagnostic and surveillance activities are vital to determine measures needed to control antimicrobial resistance (AMR), new and rapid laboratory methods are necessary to facilitate this important effort. DNA microarray technology allows the detection of a large number of genes in a single reaction. This technology is simple, specific and high-throughput. We have developed a bacterial antimicrobial resistance gene DNA microarray that will allow rapid antimicrobial resistance gene screening for all Gram-positive and Gram-negative bacteria. A prototype microarray was designed using a 70-mer based oligonucleotide set targeting AMR genes of Gram-negative and Gram-positive bacteria. In the present version, the microarray consists of 182 oligonucleotides corresponding to 166 different acquired AMR gene targets, covering most of the resistance genes found in both Gram-negative and -positive bacteria. A test study was performed on a collection of Staphylococcus aureus isolates from milk samples from dairy farms in Québec, Canada. The reproducibility of the hybridizations was determined, and the microarray results were compared with those obtained by phenotypic resistance tests (either MIC or Kirby-Bauer). The microarray genotyping demonstrated a correlation between penicillin, tetracycline and erythromycin resistance phenotypes with the corresponding acquired resistance genes. The hybridizations showed that the 38 antimicrobial resistant S. aureus isolates possessed at least one AMR gene.
由于诊断和监测活动对于确定控制抗微生物药物耐药性(AMR)所需的措施至关重要,因此需要新的和快速的实验室方法来促进这一重要工作。DNA 微阵列技术允许在单个反应中检测大量基因。该技术简单、特异、高通量。我们开发了一种用于检测所有革兰氏阳性和革兰氏阴性细菌的抗微生物药物耐药性基因的 DNA 微阵列。使用针对革兰氏阴性和革兰氏阳性细菌的 AMR 基因的 70 -mer 基寡核苷酸组设计了原型微阵列。在目前的版本中,微阵列由 182 个寡核苷酸组成,对应 166 个不同的获得性 AMR 基因靶标,涵盖了革兰氏阴性和阳性细菌中发现的大多数耐药基因。对来自加拿大魁北克省奶牛场牛奶样本的金黄色葡萄球菌分离株进行了测试研究。确定了杂交的重现性,并将微阵列结果与表型耐药性测试(MIC 或 Kirby-Bauer)的结果进行了比较。微阵列基因分型表明青霉素、四环素和红霉素耐药表型与相应的获得性耐药基因之间存在相关性。杂交表明 38 株抗微生物药物耐药的金黄色葡萄球菌分离株至少携带一个 AMR 基因。