Leptospira spp.

This volume looks at all aspects of manipulation of Leptospira spp. from strain isolation to the latest techniques used to study the pathogenesis of leptospirosis. The chapters in this book cover topics such as the procedure to cultivate and isolate leptospires from both clinical and environmental samples; using methods like whole genome sequencing and Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry to identify bacterial species; tools for gene inactivation and in vitro and in vivo assays to study the pathogenesis of leptospirosis; and the use of hamsters to evaluate leptospiral virulence and vaccine candidates. 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 comprehensive, Leptospira spp.: Methods and Protocols is a valuable resource for researchers interested in learning more about this developing field and these fascinating organisms.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Klappentext

This volume looks at all aspects of manipulation of Leptospira spp. from strain isolation to the latest techniques used to study the pathogenesis of leptospirosis. The chapters in this book cover topics such as the procedure to cultivate and isolate leptospires from both clinical and environmental samples; using methods like whole genome sequencing and Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry to identify bacterial species; tools for gene inactivation and in vitro and in vivo assays to study the pathogenesis of leptospirosis; and the use of hamsters to evaluate leptospiral virulence and vaccine candidates. 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 comprehensive, Leptospira spp.: Methods and Protocols is a valuable resource for researchers interested in learning more about this developing field and these fascinating organisms.

Zusammenfassung

Inhalt
Isolation and Culture of Leptospira from Clinical and Environmental Samples.- Core Genome Multi-Locus Sequence Typing Analyses of Leptospira spp. using the Bacterial Isolate Genome Sequence Database.- Use of MALDI-ToF Mass Spectrometry for Identification of Leptospira.- Cell Enumeration of Leptospira by Flow Cytometry.- The Single-Step Method of RNA Purification Applied to Leptospira.- Purification of LPS from Leptospira.- Isolation, Purification, and Characterization of Leptophages.- Creating a Library of Random Transposon Mutants in Leptospira.- Transposon Sequencing in Leptospira interrogans.- Specific Gene Silencing in Leptospira biflexa by RNA-Guided Catalytically inactive Cas9 (dCas9).- Leptospira spp. Toolbox for Chemotaxis Assay.- In Situ Structural Analysis of Leptospira spp. by Electron Cryotomography.- Measurement of the Cell-Body Rotation of Leptospira.- In Vivo Imaging of Bioluminescent Leptospires.- Cell Monolayer Translocation Assay.- Cell Adhesion Assay to Study Leptospiral Proteins: An Approach to Investigate Host-Pathogen Interaction.- Complement Resistance Assays.- Evaluation of Intracellular Trafficking in Macrophages.- Biofilm Formation and Quantification using the 96-Microtiter Plate.- Survival Tests for Leptospira spp..- Cultivation of Leptospira interrogans within Rat Peritoneal Dialysis Membrane Chambers.- Use of Golden Syrian Hamster as an Animal Model to Study Leptospirosis-Associated Immune Responses.- Evaluation of Vaccine Candidates against Leptospirosis using Golden Syrian Hamsters.- Leptospira and Leptospirosis.- Laboratory Diagnosis of Leptospirosis.