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JAC Advance Access published online on December 19, 2003
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkh040
© 2003 by The British Society for Antimicrobial Chemotherapy
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© 2003 The British Society for Antimicrobial Chemotherapy

Original article

Drug resistance genes and trailing growth in Candida albicans isolates

Mi Kyung Lee 1 , Laura E. Williams 1 , David W. Warnock 1 , and Beth A. Arthington-Skaggs 1 *

1 Mycotic Diseases Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, N.E., Mailstop G-11, Atlanta, Georgia 30333, USA

* Corresponding author. E-mail: BSkaggs{at}cdc.gov.

Received 17 June 2003 ; revised 10 October 2003 ; accepted 29 October 2003

Abstract

Objectives: To investigate possible molecular mechanisms of azole resistance among fluconazole-susceptible bloodstream isolates of Candida albicans that displayed the trailing growth phenomenon, and to compare these isolates with bloodstream and mucosal isolates that showed reduced susceptibilities to fluconazole.

Methods: Twelve C. albicans isolates--seven trailing and five susceptible dose dependent (SDD) or resistant (R)--were screened for ERG11 mutations by DNA sequencing and quantification of ERG11, CDR1 and MDR1 expression by RT-PCR using the LightCycler high-speed PCR system.

Results: SDD and R isolates possessed more homozygous ERG11 mutations than did the trailing isolates. Two of these, V404I and V509M, have not been described previously and were found exclusively in fluconazole SDD and R isolates. Quantification of ERG11 expression revealed that both trailing and SDD and R isolates were capable of ERG11 up-regulation in response to fluconazole, although the SDD and R isolates showed maximal up-regulation at higher fluconazole concentrations. Quantification of CDR1 and MDR1 revealed that all isolates, regardless of in vitro fluconazole response, were capable of CDR1 and MDR1 up-regulation following fluconazole exposure. Furthermore, the SDD and R isolates expressed higher constitutive levels of CDR1 and MDR1 or CDR1, respectively, in the absence of drug compared with trailing isolates.

Conclusions: Trailing isolates, although susceptible to fluconazole, express the same molecular mechanisms as SDD and R isolates following fluconazole exposure but regulate them differently.

Keywords: C. albicans, azole drug resistance, molecular mechanisms
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