Rapid Detection of Triazole Antifungal Resistance in
            <i>Aspergillus fumigatus</i>

García‐Effrón, Guillermo; Dilger, Amanda; Alcàzar-Fuoli, Laura; Park, Steven; Mellado, Emilia; Perlin, David S.

doi:10.1128/jcm.02330-07

Cited by 93 publications

(65 citation statements)

References 44 publications

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“…Other limitations of this methodology are its inability to detect newly described mutations or other potential non-FKS-linked mechanisms associated with echinocandin resistance and the possibility of changes in epidemiology making the detection of the described mutations useless. However, these potential drawbacks are shared with any molecular method designed for the detection of mechanisms of resistance (32)(33)(34). Nevertheless, this methodology is suitable to be modified by adding or eliminating PCRs in order to adapt it to detect emerging mechanisms of resistance.…”

Section: Discussionmentioning

confidence: 99%

Set of Classical PCRs for Detection of Mutations in Candida glabrata FKS Genes Linked with Echinocandin Resistance

et al. 2014

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Clinical echinocandin resistance among Candida glabrata strains is increasing, especially in the United States. Antifungal susceptibility testing is considered mandatory to guide therapeutic decisions. However, these methodologies are not routinely performed in the hospital setting due to their complexity and the time needed to obtain reliable results. Echinocandin failure in C. glabrata is linked exclusively to Fks1p and Fks2p amino acid substitutions, and detection of such substitutions would serve as a surrogate marker to identify resistant isolates. In this work, we report an inexpensive, simple, and quick classical PCR set able to objectively detect the most common mechanisms of echinocandin resistance in C. glabrata within 4 h. The usefulness of this assay was assessed using a blind collection of 50 C. glabrata strains, including 16 FKS1 and/or FKS2 mutants.

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Section: Discussionmentioning

confidence: 99%

Set of Classical PCRs for Detection of Mutations in Candida glabrata FKS Genes Linked with Echinocandin Resistance

et al. 2014

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“…The predominant resistance mechanism involves mutations in the cyp51A gene, which encodes a protein targeted by azole antifungal drugs, and a variety of these mutations have been shown to confer azole resistance (57). Molecular resistance testing has been applied to cultured isolates as a part of epidemiologic drug resistance surveillance studies (58) and to clinical isolates as a supplement to phenotypic susceptibility testing (59). Multiplex real-time PCR has also been used to detect azole-resistant strains directly in BAL fluid from patients at risk for IA (17).…”

Section: Molecular Methods For Detection Of Antifungal Drug Resistancementioning

confidence: 99%

Molecular Diagnostic Testing for Aspergillus

Powers-Fletcher

Hanson

2016

J Clin Microbiol

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The direct detection of Aspergillus nucleic acid in clinical specimens has the potential to improve the diagnosis of aspergillosis by offering more rapid and sensitive identification of invasive infections than is possible with traditional techniques, such as culture or histopathology. Molecular tests for Aspergillus have been limited historically by lack of standardization and variable sensitivities and specificities. Recent efforts have been directed at addressing these limitations and optimizing assay performance using a variety of specimen types. This review provides a summary of standardization efforts and outlines the complexities of molecular testing for Aspergillus in clinical mycology.

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“…MBs were purchased from Biosearch Technologies, Inc. (Novato, CA). MB thermal denaturation profiling and target primer design was done as previously described (8). Real-time PCR primers were designed by using the oligonucleotide design tool of the IDT SciTools software suite (Integrated DNA Technologies, Coralville, IA) and were purchased from Integrated DNA Technologies.…”

Section: Strainsmentioning

confidence: 99%

Assessment of Two New Molecular Methods for Identification of Candida parapsilosis Sensu Lato Species

García‐Effrón¹,

Cantón²,

Pemán³

et al. 2011

J Clin Microbiol

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Candida parapsilosis sensu stricto, C. orthopsilosis, and C. metapsilosis replaced C. parapsilosis groups I, II, and III in 2005. Since then, an increased interest in studying their epidemiology has arisen based on the observed differences in antifungal susceptibilities and virulence the three species. A strict differentiation of these species cannot be achieved by phenotypic methods. We evaluate two new molecular methodologies to differentiate among these species by the use of a collection of 293 bloodstream infection isolates of C. parapsilosis sensu lato. For the first method, the isolates were studied using PCR amplification of a fragment of the C. parapsilosis sensu lato FKS1 gene and a universal primer pair followed by EcoRI enzyme digestion. The other method used the allele discrimination ability of molecular beacons in a multiplex real-time PCR format. Both methods of identification showed 100% concordance with internal transcribed spacer 1 (ITS1)/ITS2 sequencing and proved to be effective for clinical applications, even with mixed-species DNAs.

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Rapid Detection of Triazole Antifungal Resistance in Aspergillus fumigatus

Cited by 93 publications

References 44 publications

Set of Classical PCRs for Detection of Mutations in Candida glabrata FKS Genes Linked with Echinocandin Resistance

Set of Classical PCRs for Detection of Mutations in Candida glabrata FKS Genes Linked with Echinocandin Resistance

Molecular Diagnostic Testing for Aspergillus

Assessment of Two New Molecular Methods for Identification of Candida parapsilosis Sensu Lato Species

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