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This volume explores the latest techniques used to study Mycobacterium ulcerans, and more specifically M. ulcerans disease (Buruli ulcer). The chapters in this book are organized into three parts and cover methods for the detection of M. ulcerans and the analysis of host-pathogen interaction; the quantification and characterization of mycolactone, the macrolide toxin of M. ulcerans; and drug development against M. ulcerans. 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.
Authoritative and cutting-edge, Mycobacterium ulcerans: Methods and Protocols is a valuable resource that helps scientists advance their research on Buruli ulcer, which is still an under-researched field in infection biology.
Inhaltsverzeichnis
Overview: Mycobacterium ulcerans Disease (Buruli ulcer).- Immunohistochemistry: A Method to Analyze M.ulcerans Infected Skin Tissue.- Primary Isolation of Mycobacterium ulcerans.- Investigation of Mycobacterium ulcerans Glycan Interactions Using Glycan Array and Surface Plasmon Resonance.- Purification and Characterization of Extracellular Vesicles from Mycobacterium ulcerans Culture.- DNA Extraction from Clinical Specimens for the Direct Detection of Mycobacterium ulcerans by Real-Time PCR.- Extraction of DNA from Environmental Samples for the Real-Time PCR Detection of Mycobacterium ulcerans.- Detection of Mycobacterium ulcerans DNA using Real-Time PCR.- Determining Viability of M. ulcerans by 16S rRNA RT Reverse Transcriptase Real-Time PCR.- Methods and Approaches for Buruli ulcer Surveillance in Africa: Lessons Learnt and Future Directions.- Overview: Mycolactone, The Macrolide Toxin of Mycobacterium ulcerans.- Conditionsfor Handling and Optimal Storage of Mycolactone.- Mycolactone Purification from M. ulcerans Cultures and HPLC-Based Approaches for Mycolactone Quantification in Biological Samples.- Detection of Mycolactone by Thin Layer Chromatography.- Competitive ELISA for the Detection and Quantification of Mycobacterium ulcerans Mycolactone.- Biochemical and Biological Assays of Mycolactone-Mediated Inhibition of Sec61.- Overview: Development of Drugs against Mycobacterium ulcerans.- In Vitro Assessment of Drug Activity against Mycobacterium ulcerans.- Drug Efficacy Testing in the Mouse Footpad Model of Buruli ulcer.- Turbidity-Based MIC Assay and Characterization of Spontaneous Drug Resistant Mutants in Mycobacterium ulcerans.- Determination of Bioenergetic Parameters in Mycobacterium ulcerans.
