DNA microarray for detection of antibiotic resistance determinants in Bacillus anthracis and closely related Bacillus cereus.
We developed a multiplex PCR for amplification of ten genes involved in resistance to ciprofloxacin, doxycycline, rifampin, and vancomycin in Bacillus anthracis and closely related Bacillus cereus. Enzymatic labelling of PCR products followed by hybridization to oligonucleotide probes on a DNA microarray enabled simultaneous detection of resistance genes tetK, tetL, tetM, tetO, vanA, and vanB and resistance-mediating point mutations in genes gyrA, gyrB, parC, and rpoB. The presented assay allows detection of clinically relevant antibiotic resistance determinants within 4h and can be used as a time-saving tool supporting conventional culture-based diagnostics.
Antwerpen
Markus H
MH
Robert Koch-Institut, Burgstrasse 37, 38855 Wernigerode, Germany.
Schellhase
Michaela
M
Ehrentreich-Förster
Eva
E
Bier
Frank
F
Witte
Wolfgang
W
Nübel
Ulrich
U
D001408:Bacillus anthracis / Q000235:genetics*; D001409:Bacillus cereus / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D013293:Enterococcus faecalis / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D013211:Staphylococcus aureus / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial