Preparation and application of microarrays for the detection of antibiotic resistance genes in samples isolated from Changchun, China.
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.
Lu
Hao
H
College of Animal Science and Veterinary Medicine, Jilin University, Changchun, Jilin, China.
Wang
Xinglong
X
Lang
Xulong
X
Wang
Yingchao
Y
Dang
Yuan
Y
Zhang
Fuxian
F
Tang
Jie
J
Li
Xiaoyan
X
Feng
Xin
X
D000668:Ampicillin Resistance / Q000187:drug effects / Q000235:genetics; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification*; D002681:China; D024901:Drug Resistance, Multiple, Bacterial / Q000187:drug effects* / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics*; D006801:Humans; D008826:Microbial Sensitivity Tests; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D013753:Tetracycline Resistance / Q000187:drug effects / Q000235:genetics
D000900:Anti-Bacterial Agents