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This second edition volume expands on the previous edition with new chapters discussing the latest protocols that encompass the detection, characterization, and genomic analysis of Cryptosporidium. The first few chapters in this book focus on diagnostic assays, such as traditional microscopy-based techniques, nucleic acid-based molecular detection, and immunoassay-based methodologies. Several chapters cover whole genome sequencing methodologies, genotyping approaches, and novel techniques such as hybridization bait capture analyses. A significant portion of this book is dedicated to functional genomics and experimental models, with discussions on tools used for genetic manipulation (including CRISPR/cas9), gene silencing, and host-pathogen interactions. The volume concludes with chapters exploring drug discovery and testing, highlighting high-content screening methods, phenotypic assays, and efficacy testing of potential disinfectants and therapeutics. 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, Cryptosporidium: Methods and Protocols, Second Edition is a valuable tool for researchers interested in gaining more insight on foundational and innovative Cryptosporidium research, which will inspire future advancements in this field.
List of contents
Cryptosporidium Diagnostic Assays: Microscopy.- Cryptosporidium Diagnostic Assays: Molecular Detection and Species Determination by Real-Time PCR-Based Assays.- Detection of Cryptosporidium Recovered from Large-Volume Water Samples Using Dead-End Ultrafiltration.- Genomic Characterization of Cryptosporidium spp. via iNextEra Library Preparation and Hybridization Bait Capture.- Preparation of Cryptosporidium for Whole Genome Sequencing.- A Real-Time PCR Based Method for the Sequence Determination of Cryptosporidium hominis and Cryptosporidium parvum gp60 Subtypes During Epidemiological Investigations.- Cryptosporidium Genotyping for Epidemiology Tracking.- A Multi-locus Variable Number Tandem Repeat Analysis (MLVA) Scheme for Epidemiological Investigations of Cryptosporidium parvum.- Optimized Genome Assembly Protocol for Cryptosporidium.- Evolutionary Genomic Analyses of Cryptosporidium.- A Method to Silence Genes in Cryptosporidium.- Use of ncRNAs to Study Host Cell-Parasite Interactions.- Genetic Manipulation of Cryptosporidium parvum.- Mouse Models for Use in Cryptosporidium Infection Studies, Quantification of Parasite Burden Using Flow Cytometry, qPCR and Histopathology, and Confocal Imaging of Oocysts.- Calf Clinical Model of Cryptosporidiosis for Efficacy Evaluation of Therapeutics.- Cell Culture Infectivity to Assess Chlorine Disinfection of Cryptosporidium Oocysts in Water.- High-Content Screening for Cryptosporidium Drug Discovery.- Phenotypic High-Throughput Screening and Drug Efficacy Test Against the Growth of
Summary
This second edition volume expands on the previous edition with new chapters discussing the latest protocols that encompass the detection, characterization, and genomic analysis of
Cryptosporidium.
The first few chapters in this book focus on diagnostic assays, such as traditional microscopy-based techniques, nucleic acid-based molecular detection, and immunoassay-based methodologies. Several chapters cover whole genome sequencing methodologies, genotyping approaches, and novel techniques such as hybridization bait capture analyses. A significant portion of this book is dedicated to functional genomics and experimental models, with discussions on tools used for genetic manipulation (including CRISPR/cas9), gene silencing, and host-pathogen interactions. The volume concludes with chapters exploring drug discovery and testing, highlighting high-content screening methods, phenotypic assays, and efficacy testing of potential disinfectants and therapeutics. 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,
Cryptosporidium: Methods and Protocols, Second Edition
is a valuable tool for researchers interested in gaining more insight on foundational and innovative
Cryptosporidium
research, which will inspire future advancements in this field.
