First molecular method for discriminating between Candida africana, Candida albicans, and Candida dubliniensis by using hwp1 gene

Romeo, Orazio; Criseo, Giuseppe

doi:10.1016/j.diagmicrobio.2008.05.014

Cited by 111 publications

(124 citation statements)

References 18 publications

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“…Molecular techniques offer unambiguous differentiation, and PCR-based methods are mostly employed. However, some PCR-based methods are complex and time-consuming while others are based on intronic sequences that may lead to erroneous results due to variations in intronic sequences among different strains [22][23][24]. In this study, we describe a duplex real-time PCR assay using primers derived from highly conserved rDNA sequences and used melting point analysis with SYBR Green dye for rapid detection and differentiation of strains of C. albicans and C. dubliniensis.…”

Section: Introductionmentioning

confidence: 99%

Rapid and Accurate Identification of Candida albicans and Candida dubliniensis by Real-Time PCR and Melting Curve Analysis

Asadzadeh¹,

Ahmad²,

Al‐Sweih³

et al. 2018

Med Princ Pract

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Objective: Candida albicans and Candida dubliniensis are germ tube-positive pathogenic yeast species. Accurate identification of these two species is warranted since C. albicans is a highly pathogenic species while C. dubliniensis exhibits increased adherence to buccal epithelial cells, reduced susceptibility to azoles and resistance to flucytosine. We have developed a duplex real-time PCR assay for rapid detection and differentiation between clinical C. albicans and C. dubliniensis isolates. Materials and Methods: A duplex real-time PCR assay was developed by using two species-specific primer pairs and SYBR Green dye to differentiate C. albicans and C. dubliniensis isolates via melting curve analysis of real-time PCR amplicons. Amplification products were also analyzed by agarose gel electrophoresis to confirm real-time PCR results. Results: Melting temperatures (T_m) for reference strains of C. albicans and C. dubliniensis were 86.55 and 82.75°C, respectively. No amplicon was obtained with DNA from reference strains of 8 other common Candida spp. When real-time PCR was applied on 226 clinical isolates previously identified by the Vitek 2 system and/or PCR sequencing of rDNA, T_m values for C. albicans (n = 113) and C. dubliniensis (n = 98) were 86.68 ± 0.529 and 82.616 ± 0.535°C, respectively. The results were confirmed by agarose gel electrophoresis. No amplicon was obtained from 15 isolates belonging to 9 other Candida spp. Conclusions: The real-time PCR assay described here does not require prior identification of clinical yeast isolates as C. albicans/C. dubliniensis by germ tube formation and accurately reports results within 2 h. Detection of amplicons by agarose gel electrophoresis is also suitable for resource-poor settings devoid of real-time PCR facilities.

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Section: Introductionmentioning

confidence: 99%

Rapid and Accurate Identification of Candida albicans and Candida dubliniensis by Real-Time PCR and Melting Curve Analysis

Asadzadeh¹,

Ahmad²,

Al‐Sweih³

et al. 2018

Med Princ Pract

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“…Molecular identification was performed according to methods described previously by Romeo and Criseo (16).…”

mentioning

confidence: 99%

Molecular Epidemiology of Candida albicans and Its Closely Related Yeasts Candida dubliniensis and Candida africana

Romeo

Criseo

2009

J Clin Microbiol

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We performed a molecular study to determine the occurrence of Candida albicans, Candida africana, and Candida dubliniensis in different clinical samples. The study provides new insights into the epidemiology of candidiasis in hospitalized patients in three hospitals in southern Italy. It also reports the first detailed epidemiological data concerning the occurrence of C. africana in clinical samples.

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“…Transcriptional Profile of Biofilm-Associated Cells To explore the mechanism of FC for inhibiting biofilm formation, the selected genes such as ALS1, ALS3, HWP1, EFG1, HYR1, CPH1 and BCR1 [27][28][29][30][31][32] (Table S1) of biofilm-associated C. albicans cells and planktonic cells were measured by real-time PCR. Biofilm-associated adherent cells were cultured with drugs (fluconazole at the concentration of 150 ng/mL, AmB at the concentration of 10 µg/mL, and FC at the concentrations of 20, 10 µg/mL) or without drugs.…”

Section: Inhibition Of C Albicans Biofilm Formation With Fcmentioning

confidence: 99%

The Activity of Fungichromin against the Formation of Candida albicans Biofilm

Wang

et al. 2016

Biological & Pharmaceutical Bulletin

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The effect of fungichromin (FC) on the formation of Candida albicans biofilm was assessed using 2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction method, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Results showed that FC revealed an inhibitory effect on the formation of C. albicans biofilm in a dose-dependent manner with a minimum inhibitory concentration (MIC) of 10 µg/mL. Over 80% of biofilm formation was inhibited by FC at the concentration of 40 µg/mL when compared with the control. Similarly, real-time PCR showed that the expression of the genes such as ALS1, ALS3, HWP1, EFG1, HYR1, CPH1 and BCR1 appeared to be remarkably affected by FC at the concentration of 20 µg/mL during the biofilm formation. In addition, FC could also induce the apoptosis of C. albicans cells in a dose-dependent manner. Therefore, FC displayed potent activity against the formation of C. albicans biofilm in vitro and played an important role in reducing the incidence of device-associated infections. Key words fungichromin (FC); Candida albicans; biofilmCandida albicans is a dimorphic yeast that can be either a commensal or an opportunistic pathogen with the capability to cause a variety of infections, ranging from superficial mucous membrane infection to life-threatening systemic diseases. 1,2)Invasive candidiasis carries unacceptably high mortality, which is reported to be as high as 40-60%.3) One of the major reasons for this phenomenon is that candidiasis can adapt to a variety of environmental conditions through attaching to the surface and growing in microbial community in a mode of biofilm.4) Biofilm can be formed not only on host tissues, but also on prosthesis and indwelling medical devices, such as urinary and vascular catheters, (dental) implants, prostheses, and heart valves.5,6) C. albicans biofilm can lead to a significantly increased resistance to conventional antifungal drugs. 7)Another important reason for the low success rate in treating candidiasis is due to the limited number of antifungal agents. Also, the treatment efficacy of antifungal agents is limited because of toxicity and resistance. 8,9) Fungichromin (FC), as one of polyene antibiotics, has been reported for its in vitro antifungal properties including C. albicans [10][11][12] ; however, until now, the activity of FC against sessile cells in C. albicans biofilm has never been studied. The aim of the present study is to explore the inhibitory effect of FC on the formation of C. albicans biofilm so as to evaluate the potential therapeutic implication for biofilm-associated candidal infections. MATERIALS AND METHODS Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC)C. albicans was prepared using broth dilution technique. FC with the gradient concentration of 0.625-80 µg/mL was prepared by two-fold serial dilution technique through RPMI 1640 medium buffered to pH 7.0 with MOPS buffer, and then inoculated with 100 µL of C. albicans suspension. Fluconazole...

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First molecular method for discriminating between Candida africana, Candida albicans, and Candida dubliniensis by using hwp1 gene

Cited by 111 publications

References 18 publications

Rapid and Accurate Identification of <b><i>Candida albicans</i></b> and <b><i>Candida dubliniensis</i></b> by Real-Time PCR and Melting Curve Analysis

Rapid and Accurate Identification of <b><i>Candida albicans</i></b> and <b><i>Candida dubliniensis</i></b> by Real-Time PCR and Melting Curve Analysis

Molecular Epidemiology of Candida albicans and Its Closely Related Yeasts Candida dubliniensis and Candida africana

The Activity of Fungichromin against the Formation of <i>Candida albicans</i> Biofilm

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