Microsatellite Genotyping Clarified Conspicuous Accumulation of Candida parapsilosis at a Cardiothoracic Surgery Intensive Care Unit

Diab-Elschahawi, Magda; Forstner, Christina; Hagen, Ferry; Meis, Jacques F.; Lassnig, Andrea; Presterl, Elisabeth; Klaassen, Corné H. W.

doi:10.1128/jcm.01179-12

Cited by 48 publications

(50 citation statements)

References 24 publications

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“…Microsatellite profiles were imported into BioNumerics v6.6 (Applied Maths, Sint-Martens-Latem, Belgium), and a dendrogram was generated by treating the data as categorical values, followed by cluster analysis by the unweighted-pair group method using average linkages. A comparison was made with a selection of Austrian isolates from a recent study (34).…”

Section: Methodsmentioning

confidence: 99%

“…Genomic DNA of C. parapsilosis isolates was extracted from 48-h-old cultures with the MagNA Pure 96 platform (Roche Diagnostics, Almere, The Netherlands) as described previously (33). Two multiplex PCRs were performed to amplify three trinucleotide repeat regions in one PCR and three hexanucleotide repeat regions in the second PCR; the setup of these assays has been described previously (34). Subsequently, the PCR products were diluted 20ϫ with double-distilled H 2 O (ddH 2 O) and 1.0 l of this dilution was added to a mixture of 0.1 l of CC-500ROX (Promega, Leiden, The Netherlands) and 8.9 l of ddH 2 O.…”

Section: Methodsmentioning

confidence: 99%

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Candida parapsilosis Resistance to Fluconazole: Molecular Mechanisms and In Vivo Impact in Infected Galleria mellonella Larvae

Souza

Fuchs

Pinhati

et al. 2015

Antimicrob Agents Chemother

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113

103

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f Candida parapsilosis is the main non-albicans Candida species isolated from patients in Latin America. Mutations in the ERG11 gene and overexpression of membrane transporter proteins have been linked to fluconazole resistance. The aim of this study was to evaluate the molecular mechanisms in fluconazole-resistant strains of C. parapsilosis isolated from critically ill patients. The identities of the nine collected C. parapsilosis isolates at the species level were confirmed through molecular identification with a TaqMan qPCR assay. The clonal origin of the strains was checked by microsatellite typing. The Galleria mellonella infection model was used to confirm in vitro resistance. We assessed the presence of ERG11 mutations, as well as the expression of ERG11 and two additional genes that contribute to antifungal resistance (CDR1 and MDR1), by using real-time quantitative PCR. All of the C. parapsilosis (sensu stricto) isolates tested exhibited fluconazole MICs between 8 and 16 g/ml. The in vitro data were confirmed by the failure of fluconazole in the treatment of G. mellonella infected with fluconazole-resistant strains of C. parapsilosis. Sequencing of the ERG11 gene revealed a common mutation leading to a Y132F amino acid substitution in all of the isolates, a finding consistent with their clonal origin. After fluconazole exposure, overexpression was noted for ERG11, CDR1, and MDR1 in 9/9, 9/9, and 2/9 strains, respectively. We demonstrated that a combination of molecular mechanisms, including the presence of point mutations in the ERG11 gene, overexpression of ERG11, and genes encoding efflux pumps, are involved in fluconazole resistance in C. parapsilosis.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Candida parapsilosis Resistance to Fluconazole: Molecular Mechanisms and In Vivo Impact in Infected Galleria mellonella Larvae

Souza

Fuchs

Pinhati

et al. 2015

Antimicrob Agents Chemother

Self Cite

113

103

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“…Multilocus sequence typing (MLST) was performed as previously described for the six major species C. albicans (23), C. glabrata (24), C. krusei (25), Candida tropicalis (26), Candida dubliniensis (27), and S. cerevisiae (28), and microsatellite typing was used for C. parapsilosis (29). MLST profiles were assigned according to corresponding pubMLST databases, when available, or assigned as novel for S. cerevisiae, starting from "1" for each allele.…”

Section: Figmentioning

confidence: 99%

Posttreatment Antifungal Resistance among Colonizing Candida Isolates in Candidemia Patients: Results from a Systematic Multicenter Study

Jensen

Johansen

Søes

et al. 2016

Antimicrob Agents Chemother

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The prevalence of intrinsic and acquired resistance among colonizing Candida isolates from patients after candidemia was investigated systematically in a 1-year nationwide study. Patients were treated at the discretion of the treating physician. Oral swabs were obtained after treatment. Species distributions and MIC data were investigated for blood and posttreatment oral isolates from patients exposed to either azoles or echinocandins for <7 or >7 days. Species identification was confirmed using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and internal transcribed spacer (ITS) sequencing, susceptibility was examined by EUCAST EDef 7.2 methodology, echinocandin resistance was examined by FKS sequencing, and genetic relatedness was examined by multilocus sequence typing (MLST). One hundred ninety-three episodes provided 205 blood and 220 oral isolates. MLST analysis demonstrated a genetic relationship for 90% of all paired blood and oral isolates. Patients exposed to azoles for >7 days (n ‫؍‬ 93) had a significantly larger proportion of species intrinsically less susceptible to azoles (particularly Candida glabrata) among oral isolates than among initial blood isolates (36.6% versus 12.9%; P < 0.001). A similar shift toward species less susceptible to echinocandins among 85 patients exposed to echinocandins for >7 days was not observed (4.8% of oral isolates versus 3.2% of blood isolates; P > 0.5). Acquired resistance in Candida albicans was rare (<5%). However, acquired resistance to fluconazole (29.4%; P < 0.05) and anidulafungin (21.6%; P < 0.05) was common in C. glabrata isolates from patients exposed to either azoles or echinocandins. Our findings suggest that the colonizing mucosal microbiota may be an unrecognized reservoir of resistant Candida species, especially C. glabrata, following treatment for candidemia. The resistance rates were high, raising concern in general for patients exposed to antifungal drugs.

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“…The discrimination of C. parapsilosis strains is fundamental not only for the rapid identification of the strains involved in the infection but also to clarify nosocomial cross-transmission and possible routes of transmission in hospital settings (van Asbeck et al, 2009). Since the description of a microsatellite multiplex strategy for C. parapsilosis strain differentiation (Sabino et al, 2010), this method has been applied in several studies mainly regarding outbreaks (Diab-Elschahawi et al, 2012;Romeo et al, 2013;Vaz et al, 2011) and is described as the most discriminatory method for C. parapsilosis strain differentiation. Herein, we used this method to genotype presumed C. parapsilosis isolates involved in fungaemia episodes from three hospitals in Brazil and to determine the patterns of relatedness of the isolates, including the pair identified in the bloodstream/catheter.…”

Section: Introductionmentioning

confidence: 99%

Different scenarios for Candida parapsilosis fungaemia reveal high numbers of mixed C. parapsilosis and Candida orthopsilosis infections

Barbedo

Vaz

Pais

et al. 2015

Journal of Medical Microbiology

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Microsatellite Genotyping Clarified Conspicuous Accumulation of Candida parapsilosis at a Cardiothoracic Surgery Intensive Care Unit

Cited by 48 publications

References 24 publications

Candida parapsilosis Resistance to Fluconazole: Molecular Mechanisms and In Vivo Impact in Infected Galleria mellonella Larvae

Candida parapsilosis Resistance to Fluconazole: Molecular Mechanisms and In Vivo Impact in Infected Galleria mellonella Larvae

Posttreatment Antifungal Resistance among Colonizing Candida Isolates in Candidemia Patients: Results from a Systematic Multicenter Study

Different scenarios for Candida parapsilosis fungaemia reveal high numbers of mixed C. parapsilosis and Candida orthopsilosis infections

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