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This book encompasses broad aspects of Cryptosporidium research with established methods that have been improved and expanded over the years as well as recent cutting-edge techniques. Within this collection are numerous molecular methods as well as protocols for genotyping and diagnostics, while also including room for in vitro cell culture techniques to address the issues with growing this difficult organism continuously. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Cryptosporidium: Methods and Protocols serves as an idea guide to the inherent challenges of working with cryptosporidial parasites to provide a foundation for new investigators to build upon.
The chapter "Accessing Cryptosporidium Omic and Isolate Data via CryptoDB.org" is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
List of contents
Cryptosporidium Diagnostic Assays: Microscopy.- Cryptosporidium Diagnostic Assays: Molecular Detection.- Detection of Cryptosporidium Recovered From Large-Volume Water Samples Using Dead-End Ultrafiltration.- Cryptosporidium Oocyst Purification Using Discontinuous Gradient Centrifugation.- Measuring Cryptosporidium Serologic Responses by Multiplex Bead Assay.- Production and Purification of Functional Cryptosporidium Glycoproteins by Heterologous Expression in Toxoplasma gondii.- Cryptosporidium Genotyping for Epidemiology Tracking.- Use of Markers to Determine Cryptosporidium Genotypes for Epidemiology Tracking and Detection.- Preparation of Cryptosporidium DNA for Whole Genome Sequencing.- Accessing Cryptosporidium Omic and Isolate Data Via CryptoDB.org.- A Novel Method to Silence Genes in Cryptosporidium.- Use of miRNAs to Study Host Cell-Parasite Interactions.- Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9.- Mouse Models for Use in Cryptosporidium Infection Studies and Quantification of Parasite Burden Using Flow Cytometry, qPCR, and Histopathology.- 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.- High-Throughput Screening of Drugs Against the Growth of Cryptosporidium parvum In Vitro by qRT-PCR.- In Vitro Culture of Cryptosporidium parvum Using Hollow Fiber Bioreactor: Applications for Simultaneous Pharmacokinetic and Pharmacodynamic Evaluation of Test Compounds.- In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers.- Two and Three Dimensional Bioengineered Human Intestinal Tissue Models for Cryptosporidium.
Summary
This book encompasses broad aspects of Cryptosporidium research with established methods that have been improved and expanded over the years as well as recent cutting-edge techniques. Within this collection are numerous molecular methods as well as protocols for genotyping and diagnostics, while also including room for in vitro cell culture techniques to address the issues with growing this difficult organism continuously. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Cryptosporidium: Methods and Protocols serves as an idea guide to the inherent challenges of working with cryptosporidial parasites to provide a foundation for new investigators to build upon.
The chapter “Accessing Cryptosporidium Omic and Isolate Data via CryptoDB.org” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
