Expression Pattern of Drug-Resistance Genes in Candida Albicans at Different Fluconazole Concentrations
Abstract
Candida albicans is an opportunistic fungus that is part of the normal flora of our
gastrointestinal and urinary tracts, but can cause infections in immune compromised
individuals. Fluconazole (FLC), which is a fungistatic azole, is a common drug used for
treatment of C. albicans infection by targeting lanosterol demethylase (ERG11). Some
clinical isolates show resistance to FLC, which can be attributed to an over expression of
ABC transporters (CDR1, CDR2), Major Facilitator Transporter (MDR1), and the azole
target (ERG11). Commonly the expression pattern of these genes is compared across
strains of susceptible and resistant strains in the absence of drugs. However, gene
expression levels of each strain may be different in different drug doses. This study
investigates the pattern of gene expression of MDR1, CDR1, CDR2, and ERG11 in
different C. albicans matched isolates at different doses of FLC. Along with the wildtype
strain SC5314, isolates from a single patient with varying susceptibility to FLC were
selected. The Minimum Inhibitory Concentration (MIC80) to FLC for each strain was
determined by microbroth dilution. The mRNA expression of the genes was analyzed using Quantitative Real-Time PCR (qRT-PCR) at FLC concentrations 4-fold lower, 4-
fold higher, and at their respective MICs. Gene expression was analyzed with and
without FLC induction and the MDR1/CDR1/CDR2/ERG11 gene expression levels of all
the strains were normalized to their uninduced expression levels. The experiments show
that in resistant strains, the genes MDR1 and ERG11 are over expressed even without
FLC induction, consistent with previous work. Increasing the FLC concentration does not
have any significant effect on gene expression in resistant strains. For the azolesusceptible
strains, the gene expression of CDR1 and CDR2 increases at FLC
concentration below its MIC80 and declines at concentrations above the MIC80. Hence,
the gene expressions of clinical isolates vary according to their MIC80 values.
Table of Contents
Introduction -- Materials and methods -- Results -- Discussion
Degree
M.S.