|Title:||Species identification of medically important fungi by use of real-time LightCycler PCR||Authors:||Hsu M.-C.
|Issue Date:||2003||Journal Volume:||52||Journal Issue:||12||Start page/Pages:||1071-1076||Source:||Journal of Medical Microbiology||Abstract:||
Invasive fungal infection has become a major cause of morbidity and mortality in immunocompromised patients. Rapid identification of pathogenic fungi to species level is critical for disease treatment. A real-time LightCycler assay aiming at rapid detection and species identification of pathogenic fungi from clinical isolates was developed. Template DNAs of different species were amplified and detected in real time by employing SYBR Green fluorescent dye. The target sequences for species-level detection were located between the 18S and 28S rDNA. Seven fungal species encountered frequently in the clinical setting, Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, Candida tropicalis, Candida guilliermondii and Cryptococcus neoformans, could be discriminated by species-specific primers and confirmed by melting-curve analyses. The range of linearity was from 1 ng to 1 pg (μl-1 water) and the sensitivity was 1 pg fungal DNA μl -1. Identification by this real-time PCR method matched biochemical identification for all 58 clinical strains. Therefore, the method is simple, rapid and sensitive enough for detection and identification of several fungal species.
|ISSN:||0022-2615||DOI:||10.1099/jmm.0.05302-0||SDG/Keyword:||DNA; green fluorescent protein; ribosome DNA; water; analytic method; article; Candida albicans; Candida glabrata; Candida guilliermondii; Candida krusei; Candida tropicalis; controlled study; Cryptococcus neoformans; DNA determination; DNA template; fungal detection; fungal strain; fungus; fungus isolation; gene amplification; gene sequence; gene targeting; nonhuman; nucleotide sequence; priority journal; real time polymerase chain reaction; sequence alignment; species identification; human; isolation and purification; methodology; polymerase chain reaction; sensitivity and specificity; Fungi; Humans; Polymerase Chain Reaction; Sensitivity and Specificity
|Appears in Collections:||醫學系|
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.