| Resumo: |
The current knowledge about the epidemiology of Legionella spp. is based mainly on data gathered from studies of outbreaks. The standard method for the detection of Legionella in environmental samples is culture on BCYE agar supplemented with L-cysteine. However it is very time-consuming and fraught with limitations, since it cannot detect viable but non-culturable cells (VBNC). In other hand, PCR basedmethods overestimate results as it detects both live and killed Legionella. The goal of this project is to develop a more reliable and rapid (3h) alternative ISH procedure for Legionella detection using biotinylated nucleic acids probes and an enzymatic reporter procedure. With this ColorISH method it is expected a direct and simple visualisation of the Legionella presence in water avoiding the need for sample manipulation and special
equipment for visualization. This may grant the non-progression of outbreaks, hence benefiting both the society and the business sector. |