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E-raamat: Cryptosporidium: Methods and Protocols

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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.
Preface v
Contributors ix
1 Cryptosporidium Diagnostic Assays: Microscopy 1(10)
Guy Robinson
Rachel M. Chalmers
2 Cryptosporidium Diagnostic Assays: Molecular Detection 11(12)
Guy Robinson
Kristin Elwin
Rachel M. Chalmers
3 Detection of Cryptosporidium Recovered from Large-Volume Water Samples Using Dead-End Ultrafiltration 23(20)
Amy M. Kahler
Vincent R. Hill
4 Cryptosporidium Oocyst Purification Using Discontinuous Gradient Centrifugation 43(18)
Michael J. Arrowood
5 Measuring Cryptosporidium Serologic Responses by Multiplex Bead Assay 61(26)
Jeffrey W. Priest
Delynn M. Moss
6 Production and Purification of Functional Cryptosporidium Glycoproteins by Heterologous Expression in Toxoplasma gondii 87(16)
Iwona Driskell
Roberta M. O'Connor
7 Cryptosporidium Genotyping for Epidemiology Tracking 103(14)
Dawn M. Roellig
Lihua Xiao
8 Use of Markers to Determine Cryptosporidium Genotypes for Epidemiology Tracking and Detection 117(12)
Anson V. Koehler
Jan Slapeta
9 Preparation of Cryptosporidium DNA for Whole Genome Sequencing 129(10)
Guy Robinson
Rachel M. Chalmers
10 Accessing Cryptosporidium Omic and Isolate Data via CryptoDB.org 139(54)
Susanne Warrenfeltz
Jessica C. Kissinger
11 A Novel Method to Silence Genes in Cryptosporidium 193(12)
Alejandro Castellanos-Gonzalez
12 Use of miRNAs to Study Host Cell-Parasite Interactions 205(14)
Ji-Hao Xu
Nicholas W. Mathy
Xian-Ming Chen
13 Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9 219(10)
Adam Sateriale
Mattie Pawlowic
Sumiti Vinayak
Carrie Brooks
Boris Striepen
14 Mouse Models for Use in Cryptosporidium Infection Studies and Quantification of Parasite Burden Using Flow Cytometry, qPCR, and Histopathology 229(24)
Karin Sonzogni-Desautels
Jan R. Mead
Momar Ndao
15 Calf Clinical Model of Cryptosporidiosis for Efficacy Evaluation of Therapeutics 253(30)
Michael W. Riggs
Deborah A. Schaefer
16 Cell Culture Infectivity to Assess Chlorine Disinfection of Cryptosporidium Oocysts in Water 283(20)
Jennifer L. Murphy
Michael J. Arrowood
17 High-Content Screening for Cryptosporidium Drug Discovery 303(16)
Melissa S. Love
Case W. McNamara
18 High-Throughput Screening of Drugs Against the Growth of Cryptosporidium parvum In Vitro by qRT-PCR 319(16)
Haili Zhang
Guan Zhu
19 In Vitro Culture of Cryptosporidium parvum Using Hollow Fiber Bioreactor: Applications for Simultaneous Pharmacokinetic and Pharmacodynamic Evaluation of Test Compounds 335(16)
Nigel Tarlett
Mary Morada
Mohini Gobin
Wesley Van Voorhis
Samuel Arnold
20 In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers 351(22)
Georgia Wilke
Ti Wang
Soumya Ravindran
Thaddeus Stappenbeck
William H. Witola
L. David Sibley
21 Two- and Three-Dimensional Bioengineered Human Intestinal Tissue Models for Cryptosporidium 373(30)
Daviel Cardenas
Seema Bhakbandra
Hymlaire Lamisere
Ying Chen
Xi-Lei Zeng
Sasirekha Ramani
Umesh C. Karandikar
David L. Kaplan
Mary K. Estes
Honorine D. Ward
Index 403