BeschreibungWe present here a novel approach for rapid genotypic identification of antibiotic resistance and infectious bacteria by combining PCR amplification with subsequent microarray analysis. A prototype microarray for the detection of 12 different antibiotic resistance genes was established. The diagnostic test (ABR-ARChipTM) is based on multiplex-PCR, incorporation of fluorescently labeled nucleotides (dCTP-Cy5) during a primer extension step and subsequent hybridization onto a microarray containing specific oligonucleotide probes. The ABR-ARChipTM was tested with genomic DNA from numerous clinical isolates of Staphylococcus epidermidis showing various spectra of antibiotic resistance. Results obtained by microarray analysis were compared with those from conventional disk diffusion testing. In parallel to the ABR-ARChipTM prototype, we have started to develop a pathogen identification microarray including -at this stage- oligonucleotide probes for Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium and Escherichia coli, representing the most frequent pathogens in community- and hospital-acquired infections. The efficiency and specificity of the designed microarray probes as well as various target labeling strategies were tested. From our results we conclude that DNA microarrays are useful tools for rapid detection of antibiotic resistance as well as infectious bacteria and can be adapted to any clinical scenario.
|26 Jan. 2004 → 27 Jan. 2004
|Statusseminar: Chiptechnologien/Microarrays, Hygiene, Gesundheit
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