Description
Accuracy coupled with speed is of greatest importance in infectious disease management of patients already under bad health conditions. We 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 recently started to develop a pathogen identification microarray including -at this initial stage- oligonucleotide probes for the 4 most frequent pathogens in community- and hospital-acquired infections. The procedure here includes PCR amplification of the bacterial 16S rRNA gene with universal primers and subsequent microarray hybridization of Cy3-labeled RNA target molecules (generated by in vitro transcription after PCR). 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.| Period | 23 Sept 2003 |
|---|---|
| Event title | Joint Annual Meeting 2003 |
| Event type | Other |
| Degree of Recognition | International |
Research Field
- Not defined