Investigation On Gene Analysis And Biotin Molecular Probe Of Candida Spp.

Candida spp. are the most common human opportunistic fungi which cause a broad-spectrum of pathogenic changes from superficial infections of skin and mucosa to severe internal and systemic diseases. Relatively little is known, however, of its epidemiology and pathogenesis because the investigation of strains was limited by the conventional tests for identification. In general, the current methods based on phenotypic properties are complex and cubersome, and the results are often susceptible and irreproducible. DNA probe technique has proved to be a valuable and reliable tool for microbiological research. We have applied this approach to investigate Candida spp. by extracting DNA, analyzing the restriction digest patterns of their genomes, DNA-DNA hybridization and comparing the photobiotin-labeled probes with the ³²P-labeied probe.First, the protoplasts were prepared by removing cell walls with snailase, and the cells were lysed in a buffer containing SDS. DNA was then extracted with phenol-chloroform.Second, DNA from standard strains of six Candida spp. were digested with the restriction endonuclease EcoRI. The gel electrophoresis revealed different pattterns for each of these species. Morever, two strains of C. albicans demonstrateu three intensely stanining...