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This volume covers the latest experimental protocols used by researchers to study the recently emerged fungal pathogen of humans, Candida auris, and also explores the pathogen's antifungal resistance, virulence, and pathogenicity. The chapters in this book cover topics such as the identification of C. auris in clinic samples by PCR and mass spectrometry; molecular typing to identify clade affiliation; genetic manipulation; genome analysis by bioinformatics; and phenotypic characterization using various biochemical, mass spectrometry, microscopy, and flow cytometry approaches. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and practical, Candida auris: Methods and Protocols is a valuable resource for researchers interested in learning more about this developing field.
Sommario
Isolation of Candida auris in Clinical Specimens .- Candida auris Identification and Profiling by MALDI-ToF Mass Spectrometry.- Molecular Tools for Candida auris Identification and Typing.- qPCR Detection of Candida auris using the GPS(TM) CanAur MONODOSE dtec-qPCR Test.- Susceptibility Testing of Candida auris Isolates.- Deciphering Candida auris Paradoxical Growth Effect (Eagle Effect) in Response to Echinocandins.- An Optimized Electroporation Procedure for Genetic Transformation of Candida auris.- Genetic Transformation of Candida auris via Homology-Directed Repair Using a Standard Lithium-Acetate Protocol.- Downregulation of Essential Genes in the Fungal Pathogen Candida auris.- Cell Viability Assays for Candida auris.- Measuring Gene Expression via mRNA Abundance in Candida auris.- Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Candida auris Metabolites.- Isolation of Extracellular Vesicles from Candida auris.- Labelling of Candida auris Cell Walls to Examine Levels of PAMP Exposure by Flow Cytometry and Fluorescence Microscopy.- High-Pressure Freezing and Transmission Electron Microscopy to Visualize the Ultrastructure of the C. auris Cell Wall.- Generating Complete Genome Assemblies of Candida auris.- MycoSNP: A Portable Workflow for Performing Whole-Genome Sequencing Analysis of Candida auris.- Bioinformatic Identification of ABC Transporters in Candida auris.- Examining Neutrophil-Candida auris Interactions with Human Neutrophils Ex Vivo.- Modeling Candida auris Colonization on Porcine Skin Ex Vivo.- A Human Ex Vivo Skin Model to Study Candida auris Biofilms.- In Vivo Skin Colonization/Decolonization Models for Candida auris.- Use of Galleria mellonella Larvae to Study the Pathogenicity and Clonal Lineage-Specific Behaviors of the Emerging Fungal Pathogen Candida auris.- Drosophila melanogaster as a Rapid and Reliable In Vivo Infection Model to Study the Emerging Yeast Pathogen Candida auris.- An Immunocompromised Mouse Model of Candida auris Systemic Infection.- Mouse Gastrointestinal Colonization Model for Candida auris.
