Document Detail


Mutations in the fks1 gene in Candida albicans, C. tropicalis, and C. krusei correlate with elevated caspofungin MICs uncovered in AM3 medium using the method of the European Committee on Antibiotic Susceptibility Testing.
MedLine Citation:
PMID:  18591282     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Mutations in two specific regions of the Fks1 subunit of 1,3-beta-D-glucan synthase are known to confer decreased caspofungin susceptibility on Candida spp. Clinical isolates of Candida spp. (404 Candida albicans, 62 C. tropicalis, and 21 C. krusei isolates) sent to the French National Reference Center were prospectively screened for susceptibility to caspofungin in vitro by the broth microdilution reference method of the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antibiotic Susceptibility Testing (AFST-EUCAST). Twenty-eight isolates (25 C. albicans, 2 C. tropicalis, and 1 C. krusei isolate) for which the caspofungin MIC was above the MIC that inhibited 90% of the isolates of the corresponding species (MIC(90)) were subjected to molecular analysis in order to identify mutations in the fks1 gene. Substitutions in the deduced protein sequence of Fks1 were found for 8 isolates, and 20 isolates had the wild-type sequence. Among the six C. albicans isolates harboring mutations, six patterns were observed involving amino acid changes at positions 641, 645, 649, and 1358. For C. tropicalis, one isolate showed an L644W mutation, and for one C. krusei isolate, two mutations, L658W and L701M, were found. Two media, RPMI medium and AM3, were tested for their abilities to distinguish between isolates with wild-type Fks1 and those with mutant Fks1. In RPMI medium, caspofungin MICs ranged from 0.25 to 2 microg/ml for wild-type isolates and from 1 to 8 micro for mutant isolates. A sharper difference was observed in AM3: all wild-type isolates were inhibited by 0.25 micro of caspofungin, while caspofungin MICs for all mutant isolates were >or=0.5 microg/ml. These data demonstrate that clinical isolates of C. albicans, C. tropicalis, and C. krusei with decreased susceptibility to caspofungin in vitro have diverse mutations in the fks1 gene and that AM3 is potentially a better medium than RPMI for distinguishing between mutant and wild-type isolates using the AFST-EUCAST method.
Authors:
Marie Desnos-Ollivier; Stéphane Bretagne; Dorothée Raoux; Damien Hoinard; Françoise Dromer; Eric Dannaoui;
Publication Detail:
Type:  Journal Article     Date:  2008-06-30
Journal Detail:
Title:  Antimicrobial agents and chemotherapy     Volume:  52     ISSN:  1098-6596     ISO Abbreviation:  Antimicrob. Agents Chemother.     Publication Date:  2008 Sep 
Date Detail:
Created Date:  2008-08-26     Completed Date:  2008-10-23     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0315061     Medline TA:  Antimicrob Agents Chemother     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3092-8     Citation Subset:  IM    
Affiliation:
Institut Pasteur, Centre National de Référence Mycologie et Antifongiques, Unité de Mycologie Moléculaire, CNRS URA3012, 75724 Paris Cedex 15, France.
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MeSH Terms
Descriptor/Qualifier:
Antifungal Agents / pharmacology*
Base Sequence
Candida / drug effects*,  enzymology,  genetics
Candida albicans / drug effects*,  enzymology,  genetics
Candida tropicalis / drug effects*,  enzymology,  genetics
Culture Media
Drug Resistance, Fungal
Echinocandins / pharmacology*
Europe
Fungal Proteins / genetics
Glucosyltransferases / genetics*
Humans
Microbial Sensitivity Tests / standards
Molecular Sequence Data
Mutation*
Chemical
Reg. No./Substance:
0/Antifungal Agents; 0/Culture Media; 0/Echinocandins; 0/Fungal Proteins; 0/caspofungin; EC 2.4.1.-/Glucosyltransferases; EC 2.4.1.34/1,3-beta-glucan synthase
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