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Cited by 125Antifungal resistance and susceptibilityFungal and yeast genetics researchFungal Plant Pathogen Control

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Overexpression of aldo-keto-reductase in azole-resistant clinical isolates of Candida glabrata determined by cDNA-AFLP

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Farahyar, Shirin, Zaini, Farideh, Kordbacheh, Parivash, Rezaie, Sassan, Safara, Mahin, Raoofian, Reza, Heidari, Mansour·2013·DARU Journal of Pharmaceutical Sciences
DOI 10.1186/2008-2231-21-1PMC PMC3556022
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Abstract

BACKGROUND: Candida glabrata causes significant medical problems in immunocompromised patients. Many strains of this yeast are intrinsically resistant to azole antifungal agents, and treatment is problematic, leading to high morbidity and mortality rates in immunosuppressed individuals. The primary goal of this study was to investigate the genes involved in the drug resistance of clinical isolates of C. glabrata. METHODS: The clinical isolates of C. glabrata were collected in an epidemiological survey of candidal infection in immunocompromised patients and consisted of four fluconazole and itraconazole resistant isolates, two fluconazole and itraconazole sensitive isolates, and C. glabrata CBS 138 as reference strain. Antifungal susceptibility patterns of the organisms were determined beforehand by the Clinical and Laboratory Standards Institute (CLSI). The potential gene(s) implicated in antifungal resistance were investigated using complementary DNA- Amplified Fragment Length Polymorphism (cDNA-AFLP). Semi-quantitative RT-PCR was carried out to evaluate the expression of gene(s) in resistant isolates as compared to sensitive and reference strains. RESULTS AND CONCLUSIONS: The aldo-keto-reductase superfamily (AKR gene) was upregulated in the resistant clinical isolates as assessed by cDNA-AFLP. Semi-quantitative RT-PCR revealed AKR mRNA expression approximately twice that seen in the sensitive isolates. Overexpression of the AKR gene was associated with increased fluconazole and itraconazole resistance in C. glabrata. The data suggest that upregulation of the AKR gene might give a new insight into the mechanism of azole resistance.

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Crossref · 10.1186/2008-2231-21-1

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Figures

Figure 1 Expression pattern of TDFs of cDNA-AFLP PAGE. Sensitive amplification of cDNA-AFLP on a PAGE from S3Mbo1/S4Mbo1 as primers. The lanes numbered correspond to the clinical isolates presented in Table 1. The arrows point to a differentially expressed TDFs. M: Marker 50 bp; * FLU: Fluconazole.
Figure 1 Expression pattern of TDFs of cDNA-AFLP PAGE. Sensitive amplification of cDNA-AFLP on a PAGE from S3Mbo1/S4Mbo1 as primers. The lanes numbered correspond to the clinical isolates presented in Table 1. The arrows point to a differentially expressed TDFs. M: Marker 50 bp; * FLU: Fluconazole.CC BY · PubMed Central
Figure 2 Expression pattern of TDFs of cDNA-AFLP PAGE using silver staining. Sensitive amplification of cDNA-AFLP on a PAGE from primer combinations S2Mbo1/S2Mbo1 and S2Mbo1/S3Mbo1. The lanes numbered correspond to the clinical isolates presented in Table 1. The arrows point to differentially expressed TDFs. DNA ladder; M: (100 bp) molecular weight marker. *FLU: Fluconazole.
Figure 2 Expression pattern of TDFs of cDNA-AFLP PAGE using silver staining. Sensitive amplification of cDNA-AFLP on a PAGE from primer combinations S2Mbo1/S2Mbo1 and S2Mbo1/S3Mbo1. The lanes numbered correspond to the clinical isolates presented in Table 1. The arrows point to differentially expressed TDFs. DNA ladder; M: (100 bp) molecular weight marker. *FLU: Fluconazole.CC BY · PubMed Central
Figure 3 Semi quantitative RT-PCR analysis of AKR gene mRNA expression. The lanes numbered correspond to the clinical isolates presented in Table 1. N, negative control (water). URA3 gene (173 bp) was used as internal control.
Figure 3 Semi quantitative RT-PCR analysis of AKR gene mRNA expression. The lanes numbered correspond to the clinical isolates presented in Table 1. N, negative control (water). URA3 gene (173 bp) was used as internal control.CC BY · PubMed Central
Figure 4 AKR gene showed increased expression level after normalization with internal control (URA3). Samples 1 and 2 show clinical isolates of C. glabrata sensitive to fluconazole and itraconazole; 3 shows CBS 138, and samples 4,5,6,7 are resistant clinical isolates.
Figure 4 AKR gene showed increased expression level after normalization with internal control (URA3). Samples 1 and 2 show clinical isolates of C. glabrata sensitive to fluconazole and itraconazole; 3 shows CBS 138, and samples 4,5,6,7 are resistant clinical isolates.CC BY · PubMed Central

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  1. National Committe for Clinical Laboratory Standards: NCCLS document M27-A2 National Committee for Clinical Laboratory Standards 2002. Reference method for broth dilution antifungal susceptibility testing of yeasts. 2002, Wayne, Pa: NCCLS

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Metadata

Authors
Farahyar, S., Zaini, F., Kordbacheh, P., Rezaie, S., Safara, M., Raoofian, R., & Heidari, M.
Affiliations
  • Tehran University of Medical Sciences (IR)
  • Mashhad University of Medical Sciences (IR)
Corresponding author
Zaini, Farideh
ORCID
Venue
DARU Journal of Pharmaceutical Sciences
Publisher
Springer Science and Business Media LLC
Published
2013-01-02
Volume
21
Issue
1
Pages
1-1
ISSN
  • 1560-8115 (ISSN-L)
  • 2008-2231
DOI
10.1186/2008-2231-21-1
PMC
PMC3556022
OpenAlex
W2140631543
Cited by
125
References
32
Citations by year
2026: 1 · 2023: 1 · 2022: 1 · 2021: 17 · 2020: 15 · 2019: 23 · 2018: 10 · 2017: 24 · 2016: 14 · 2015: 5 · 2014: 9 · 2013: 5
FWCI
8.02
Type
journal article
Language
en
Open access
Licence
Locations
  • DARU Journal of Pharmaceutical Sciences (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
Tehran University of Medical Sciences and Health Services
Topics
  • Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Fungal Plant Pathogen Control
Keywords
Candida glabrata, azole resistance, aldo-keto reductase, fluconazole resistance, itraconazole resistance, antifungal resistance, gene overexpression, immunocompromised patients, candidiasis, clinical isolates

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