Main description:

This detailed volume aims to elucidate the molecular mechanisms that underlie antifungal resistance. The book highlights methods to identify and characterize antifungal activity, to define and characterize strains with altered responses to antifungal drugs, to investigate the genetic and molecular mechanisms of these alterations of antifungal drug susceptibility, and, finally, to approach the study of these processes in animal models of fungal infection. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Antifungal Drug Resistance: Methods and Protocols serves as an ideal resource for the field and a guide for investigators to study this important translational aspect of fungal biology.

Contents:

Part I: Identification and Characterization of Molecules with Antifungal Activity

1. Antifungal Susceptibility of Yeasts and Filamentous Fungi by CLSI Broth Microdilution Testing

Nathan P. Wiederhold

2. Luciferase-Based High Throughput Screen with Aspergillus fumigatus to Identify Antifungal Small Molecules

Timothy J. Opperman, Sourabh Dhingra, Cecilia Gutierrez, Steven M. Kwasny, and Robert A. Cramer

3. A Dual-Readout High Throughput Screening Assay for Small Molecules Active Against Aspergillus fumigatus

Sarah R. Beattie and Damian J. Krysan

4. Antifungal Susceptibility Testing and Drug Discovery in the Dimorphic Fungus Histoplasma capsulatum

Chad A. Rappleye

5. A 384-Well Microtiter Plate Model for Candida Biofilm Formation and Its Application to High Throughput Screening

Olabayo H. Ajetunmobi, Gina Wall, Bruna V. Bonifacio, Daniel Montelongo-Jauregui, and Jose L. Lopez-Ribot

Part II: Identification and Characterization of Fungal Strains with Altered Drug Susceptibility

6. Protocols for Measuring Tolerant and Heteroresistant Drug Responses of Pathogenic Yeasts

Naomi Lyons and Judith Berman

7. Genome-Wide Identification of Variants Associated with Antifungal Drug Resistance

Aina Martinez-Zurita and Christina A. Cuomo

8. Copy Number Variation and Allele Ratio Analysis in Candida ablicans Using Whole Genome Sequencing Data

Robert T. Todd and Anna Selmecki

9. Drug Target Elucidation through Isolation and Analysis of Drug-Resistant Mutants in Cryptococcus neoformans

Michael J. Hoy and Joseph Heitman

10. Chemical-Genetic Approaches for Exploring Mode of Action of Antifungal Compounds in the Fungal Pathogen Candida albicans

Nicole Robbins, Troy Ketela, Sang Hu Kim, and Leah E. Cowen

Part III: Characterization of the Genetic and Molecular Mechanisms of Antifungal Resistance

11. Characterizing Candida glabrata Pdr1, a Hyperactive Transcription Factor Involved in Azole Resistance

Lucia Simonicova and W. Scott Moye-Rowley

12. Heterologous Expression of Candida Antifungal Target Genes in the Model Organism Saccharomyces cerevisiae

Kelley R. Healey and Ashutosh Singh

13. Conditional Protein Depletion in the Analysis of Antifungal Drug Resistance in Candida glabrata

Thomas P. Conway and W. Scott Moye-Rowley

14. Transport Across Membranes: Techniques for Measuring Efflux in Fungal Cells

Brooke D. Esquivel and Theodore C. White

15. Transport Across Membranes: Techniques for Measuring Drug Import in Fungal Cells

Brooke D. Esquivel and Theodore C. White

Part IV: Characterization of Antifungal Drug Resistance Using Animal Infection Models

16. Analysis of Candida Antifungal Resistance Using Animal Infection Models

David R. Andes and Jeniel E. Nett

17. Using Bioluminescence to Image Candida glabrata Urinary Tract Infections in Mice

Sanne Schrevens, Riccardo Torelli, Maurizio Sanguinetti, and Dominque Sanglard