DNA microarray for genotyping multidrug-resistant Pseudomonas aeruginosa clinical isolates.
The management of infections with multidrug-resistant Pseudomonas aeruginosa needs fast and reliable methods of antibiotic susceptibility testing for a therapy improvement. For this purpose, we developed a DNA microarray for genotyping antibiotic resistance and a few virulence factors. The array covers mutations in the efflux regulators mexR, nfxB, mexT, gyrase gyrA, and parC, as well as plasmid-encoded vim, imp, oxa, aph, aac, and aad genes, and virulence-associated mucA and exoU, exoT, and exoS genes, respectively. The whole procedure can be performed in less than 5 h and consists of DNA isolation, target gene amplification, fluorescence labeling, fragmentation, and array hybridization. Concerning the genotype-phenotype comparison in the test collection, the coverage of relevant resistance determinants for antibiotics used in a calculated therapy of critical ill patients was 87.8%.
Weile
Jan
J
Robert-Bosch-Hospital, Department of Laboratory Medicine, 70376 Stuttgart, Germany. jan.weile@ikp-stuttgart.de
Schmid
Rolf D
RD
Bachmann
Till T
TT
Susa
Milorad
M
Knabbe
Cornelius
C
D003428:Cross Infection / Q000145:classification / Q000188:drug therapy / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010641:Phenotype; D011552:Pseudomonas Infections / Q000145:classification / Q000188:drug therapy / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000145:classification / Q000187:drug effects / Q000235:genetics; D014774:Virulence / Q000235:genetics