Propose: to develop a new sensitive, specific and rapid method for the diagnosis of fungal eye infectious diseases, based on PCR assay. The ultimate goal is to have a rapid identification process of the pathogen which allows the rapid institution of effective antifungal therapy. We have tested the specificity and sensitivity of the method in 12 ocular fungal pathogens: five Candida species, four Aspergillus species, two Fusarium species and one Cryptococcus species. The fungal infection was induced in one eye of each of five rabbits (New Zealand) with the following pathogens: C. albicans, C. parapsilosis, A. niger, A. fumatus and F. oxysporum. The sample collection was performed during the period of infection. Cornea, aqueous and vitreous samples were taken. The detection technique involves two steps: a first PCR to amplify the 5.8S rRNA gene and the two ribosomal internal transcribed spacers (ITS) and the semi-nested PCR to improve the sensivity of the method. The DNA sequence target was detected in all infection samples studied. However, traditional microbiological techniques failed in some cultures performed, especially at the beginning of the infection. Our data suggests that the PCR amplification of 5.8S/ITS regions allow a rapid detection of fungal pathogens in eye infections.
Full conference title:
6th Congress of the European Confederation of Medical Mycology Societies
- ECMM 6th (2000)