Rapid detection and identification of Candida albicans and Torulopsis (Candida) glabrata in clinical specimens by species-specific nested PCR amplification of a cytochrome P-450 lanosterol-alpha-demethylase (L1A1) gene fragment
Details
Serval ID
serval:BIB_B71548F72852
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Rapid detection and identification of Candida albicans and Torulopsis (Candida) glabrata in clinical specimens by species-specific nested PCR amplification of a cytochrome P-450 lanosterol-alpha-demethylase (L1A1) gene fragment
Journal
Journal of Clinical Microbiology
ISSN
0095-1137
Publication state
Published
Issued date
08/1994
Peer-reviewed
Oui
Volume
32
Number
8
Pages
1902-7
Notes
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Aug
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Aug
Abstract
PCR of a Candida albicans cytochrome P-450 lanosterol-alpha-demethylase (P450-L1A1) gene segment is a rapid and sensitive method of detection in clinical specimens. This enzyme is a target for azole antifungal action. In order to directly detect and identify the clinically most important species of Candida, we cloned and sequenced 1.3-kbp fragments of the cytochrome P450-L1A1 genes from Torulopsis (Candida) glabrata and from Candida krusei. These segments were compared with the published sequences from C. albicans and Candida tropicalis. Amplimers for gene sequences highly conserved throughout the fungal kingdom were first used; positive PCR results were obtained for C. albicans, T. glabrata, C. krusei, Candida parapsilosis, C. tropicalis, Cryptococcus neoformans, and Trichosporon beigelii DNA extracts. Primers were then selected for a highly variable region of the gene, allowing the species-specific detection from purified DNA of C. albicans, T. glabrata, C. krusei, and C. tropicalis. The assay sensitivity as tested for C. albicans in seeded clinical specimens such as blood, peritoneal fluid, or urine was 10 to 20 cells per 0.1 ml. Compared with results obtained by culture, the sensitivity, specificity, and efficiency of the species-specific nested PCR tested with 80 clinical specimens were 71, 95, and 83% for C. albicans and 100, 97, and 98% for T. glabrata, respectively.
Keywords
Amino Acid Sequence
Base Sequence
Candida/enzymology/genetics/*isolation & purification
Candida albicans/enzymology/genetics/*isolation & purification
Cytochrome P-450 Enzyme System/*genetics
Genes, Fungal/*genetics
Humans
Molecular Sequence Data
Mycoses/microbiology
Oxidoreductases/*genetics
Polymerase Chain Reaction/*methods
Sensitivity and Specificity
Sequence Homology, Amino Acid
Species Specificity
Pubmed
Web of science
Create date
11/02/2008 13:39
Last modification date
20/08/2019 16:25