Amplification primers specific for yeast species and a rapid microplate assay for detection of PCR product will be developed in these studies. The B region of the large ribosomal subunit DNA (LrDNA) of 24 yeast species will be sequenced and computer alignment against known sequences will be used to select species-specific regions of LrDNA for use as PCR primer oligonucleotides. A 3-primer PCR technique (2 universal primers and one species primer) will be used to validate the usefulness of the primer for species identification. Concurrently, a microplate assay for detection of PCR product will be developed and tested using the panel of primers. The PCR primers and the microplate assay will be combined to produce assay kits for detection of fungal species. The target markets include clinical and veterinary diagnostics, industrial quality control and basic research. PROPOSED COMMERCIAL APPLICATION: Diagnostic kits for fungal identification will be developed. Target markets include clinical, veterinarian, industrial quality control and basic and applied research.
Thesaurus Terms:method development, microorganism classification, yeast Candida, Cryptococcus, Saccharomyces cerevisiae enzyme linked immunosorbent assay, nucleic acid sequence, polymerase chain reactionNational Institute of Allergy and Infectious Diseases (NIAID)
