title: Development of a high-throughput DNA microarray for drug-resistant  gene detection  and its preliminary application.
abstract: Most bacteria are resistant to a wide variety of antibiotics and other drugs, which decrease the effectiveness of clinical drug therapies. The present study developed a high-throughput DNA microarray for drug-resistant gene detection. A total of 115 specific oligonuclieotide probes with lengths of 42 nt to 45 nt and comparable Tm values were selected from 17 categories of drug-resistant genes in the National Center for Biotechnology Information database and were chemically synthesized. The entire bacterial DNA was extracted, randomly amplified, and labeled using Cy3-dCTP. The hybridization conditions of the microarray test were optimized to improve sensitivity and specificity. The drug-resistant genes were detected and genotyped using microarray analysis after hydration at 42°C for 4h with 2× hybridization solution. The microarray test sensitivity was 20ng/μL DNA. The performance of the microarray was validated using reference strains and clinical isolates. The results were consistent with direct DNA sequence analysis and drug susceptibility tests. The developed DNA microarray could be used to detect and screen drug-resistant bacteria rapidly and simultaneously. Thus, the present study could be helpful in effectively using antibiotics and controlling infectious diseases.
authors.affiliation: Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing 210029, PR China.
authors.forename: Yali
authors.initials: Y
authors.lastname: Fu
authors.forename: Ying
authors.initials: Y
authors.lastname: Pan
authors.forename: Mingjie
authors.initials: M
authors.lastname: Pan
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.forename: Wu
authors.initials: W
authors.lastname: Liu
authors.forename: Yuexi
authors.initials: Y
authors.lastname: Li
mesh_terms: D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification; D001424:Bacterial Infections / Q000382:microbiology; D019295:Computational Biology; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial; D005798:Genes, Bacterial; D005838:Genotype; D057166:High-Throughput Screening Assays / Q000379:methods; D006761:Hospitals; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D015345:Oligonucleotide Probes