DNA microarray for the detection of therapeutically relevant antibiotic resistance determinants in clinical isolates of Staphylococcus aureus.
An oligonucleotide microarray was constructed for the rapid and sensitive molecular detection of antibiotic resistance determinants in Staphylococcus aureus. The array is equipped with oligonucleotide capture probes for the detection of 10 clinically and therapeutically relevant antibiotic resistance genes and -mutations (mecA, aacA-aphD, tetK, tetM, vat(A), vat(B), vat(C), erm(A), erm(C), grlA-mutation) as well as several control probes. A microarray concept was established including multiplexed PCR amplification, DNA labeling, hybridization and data processing. This concept was applied to clinical Staphylococcus aureus isolates and results were concordant with those from standard genotypic and phenotypic resistance testing. Our microarray concept offers rapid and accurate identification of antibiotic resistance profiles. It is easily expandable and thus can be adapted to changing clinical and epidemiological requirements in clinical diagnosis as well as in epidemiological studies.
Strommenger
Birgit
B
Robert Koch Institute, Wernigerode Branch, Burgstr. 37, D-38855 Wernigerode, Germany. strommengerb@rki.de
Schmidt
Christiane
C
Werner
Guido
G
Roessle-Lorch
Beate
B
Bachmann
Till T
TT
Witte
Wolfgang
W
D015342:DNA Probes / Q000032:analysis; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D005798:Genes, Bacterial; D008826:Microbial Sensitivity Tests; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010641:Phenotype; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification*
D015342:DNA Probes