Aim: Molecular identification of pathogenic fungi
 
We are developing a reference database for molecular identification of fungi using PCR-fingerprinting and we are evaluating the roles of RFLP and gene sequence analyses of the rDNA gene cluster and the actin gene in identification. Thus far, we have tested these methods using pathogenic (and non-pathogenic) species of the genus Candida from culture collections and some clinical samples. Fingerprints have also been done on Cryptococcus neoformans and other fungal genera (e.g. Penicillium, Aspergillus, Trichoderma species).
 
 
Preliminary Assessment:
 
    * PCR fingerprinting is simple, rapid and reproducibly identifies Candida at species and strain level, including, for example, C. dubliniensis, C. norvegensis and C. inconspicua (Meyer, W et al. (1997): Electrophoresis 18; 1548-1559; Latouche, N et al. (1997): J Clin Microbiol 35; 3171-3180). Similar identification levels where obtained in C. neoformans, Penicillium and Aspergillus by this method.
    * RFLP analysis of the rDNA gene cluster has been shown to be suitable for identification to species level only.
    * Sequence analysis of the rDNA gene cluster and the actin gene is suitable for species identification but is expensive and relatively time-consuming.
 
 
Current developments:
 
    * We are evaluating our methods further in a clinical diagnostic setting and are keen to extend the number of less common Candida species and to include other fungal genera in our data-base.
    * We intend to publish the methods and examples of standard PCR-fingerprints of different fungal species on our web-site (www.usyd.edu.au/~cidm).
    * We are evaluating the direct amplification of fungal DNA from paraffin tissue for subsequent DNA sequencing.
    * Once the database is fully developed we will offer a reference facility for the tests based on cost-recovery, to complement the classical identification methods offered in Adelaide by Associate Professor David Ellis.
 
 
Proposed users and call for isolates:
 
These tests can be used for confirmation of fungal identity and in epidemiological investigations.
During development (year 2000), we invite all interested diagnostic and research labs to contribute problem fungal strains or others of special interest, including summary clinical information. We will e-mail/fax/mail results and provide contributors with regularly updated information on the status of our database.
 
Samples should be send to:
 
A/ Prof. Wieland Meyer

Molecular Mycology Research Laboratory
CIDM, ICPMR, Level 3, Room 3114A
Westmead Hospital
Darcy Road
Westmead, NSW, 2145
 
Marked: Molecular Mycology
 
Contacts:
 
Technical questions/results: A/Prof. Wieland Meyer
Phone:   61 2 9845 6895
Fax:   61 2 9891 5317
E-mail:   w.meyer@usyd.edu.au
 
Clinical questions: Dr. Sharon Chen / Prof. Tania Sorrell
Phone: 61 2 98456012