Rapid detection of point mutations by fluorescence resonance energy transfer and probe melting curves in Candida species.

摘要: Background: The LightCyclerTM combines rapid amplification of nucleic acids in glass capillaries with melting curve analysis based on fluorescence resonance energy transfer for the sensitive detection point mutations various settings, such as drug resistance and hereditary diseases. Point leading to an altered structure lanosteroldemethylase, target enzyme fungistatic azoles, are important mechanism acquired Candida albicans . Methods: We screened 13 fluconazole-resistant C. 21 tropicalis strains (minimum inhibitory concentration >128 mg/L), isolated from patients AIDS, presence defined by comparing conventional cycle sequencing a newly designed LightCycler-based assay. Results: In , 5 isolates showed wild-type sequence, 8 homozygous nucleotide exchange thymine cytosine at position 1554 (T1554C). A heterozygous genotype was detected LightCycler, but only 3 sequencing. 2 isolates, mutation amino acid 464 (glycine serine) both assays. Conclusion: LightCycler technique offers standardized, fast, sensitive, reproducible different spp.