Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Raggam, RB.
Qualitative detection of Legionella species in bronchoalveolar lavages and induced sputa by automated DNA extraction and real-time polymerase chain reaction.
[ Dissertation ] Karl Franzens Universität Graz; 2002.
- Autor*innen der Med Uni Graz:
- Raggam Reinhard Bernd
- Betreuer*innen:
- Baumann Gabriele
- Kessler Harald
- Altmetrics:
- Abstract:
- Molecular assays for qualitative detection of Legionella species (spp.) in clinical specimens were evaluated. DNA extraction was done either with a fully automated DNA extraction protocol on the MagNA Pure LC System of with manual DNA extraction. Amplification and detection were done by real-time polymerase chain reaction (PCR) on the LightCycler (LC) instrument. Oligonucleiotides were derived from the 16s rRNA gene of Legionella spp. The assays included a specially designed DNA fragment as Legionella-specific internal control (IC). For both molecular assays, the detection limit was determined to be 5 CFU per LC PCR run. Sixty-one clinical specimens were tested with the molecular assays. Results were compared to culture. Five samples were found to be positive with the molecular assays. Three of them were positive in culture. No inhibition was found throughout the whole study. In conclusion, the molecular assays described, may lead to sage and early diagnosis of Legionaires disease. They proved to be suitable for the routine molecular diagnostics laboratory.