College of Animal Science and Veterinary Medicine, Jilin University, Changchun, Jilin, China.
Mol Biol Rep. 2010 Apr;37(4):1857-65. doi: 10.1007/s11033-009-9621-4. Epub 2009 Jul 30.
The emergence of antibiotic-resistant bacteria, especially tetracycline- and beta-lactam-resistant bacteria, poses a great threat to human health. The purpose of this study was to develop and apply a suitable gene microarray for the detection of antibiotic resistance genes. We isolated 463 strains of bacteria from a hospital, a veterinary station, an animal nursery, and living environment of Changchun, China. After screening, it was found that 93.9% of these bacteria were resistant to tetracycline, 74.9% to ampicillin, 55.6% to deoxycycline, and 41.7% to ciprofloxacin. For amplification of antibiotic genes, we designed 28 pairs of primers. In addition, 28 hybridization probes for these genes were developed. The DNA microarray analysis was performed at 42 degrees C for 5 h. We were successful in detecting 12 resistance genes by microarray analysis. After detection, we also evaluated the sensitivity of the microarray analysis. The LDL (Lowest Detection Level) of the microarray was 1 x 10(6) copies/ml of template DNA. It is believed that such microarray-based determination of tetracycline and beta-lactam resistance genes can have a potential application in clinical studies in the future.
抗生素耐药菌的出现，特别是对四环素和β-内酰胺类抗生素耐药菌的出现，对人类健康构成了极大的威胁。本研究旨在开发和应用合适的基因微阵列来检测抗生素耐药基因。我们从中国长春的医院、兽医站、动物养殖场和生活环境中分离出了 463 株细菌。经过筛选，发现这些细菌中有 93.9%对四环素耐药，74.9%对氨苄西林耐药，55.6%对脱氧土霉素耐药，41.7%对环丙沙星耐药。为了扩增抗生素基因，我们设计了 28 对引物。此外，还开发了 28 个针对这些基因的杂交探针。在 42°C 下进行 DNA 微阵列分析 5 小时。我们成功地通过微阵列分析检测到 12 种耐药基因。检测后，我们还评估了微阵列分析的灵敏度。微阵列的 LDL（最低检测水平）为 1 x 10(6)拷贝/ml 模板 DNA。相信这种基于微阵列的四环素和β-内酰胺类抗生素耐药基因的测定方法在未来的临床研究中具有潜在的应用价值。
