Malaria Methods and Protocols

Front Cover
Denise L. Doolan
Springer Science & Business Media, Feb 2, 2008 - Medical - 631 pages
The Plasmodium spp. parasite was identified as the causative agent of malaria in 1880, and the mosquito was identified as the vector in 1897. Despite subsequent efforts focused on the epidemiology, cell biology, immunology, molecular biology, and clinical manifestations of malaria and the Plasmodium parasite, there is still no licensed vaccine for the prevention of malaria. Physical barriers (bed nets, window screens) and chemical prevention methods (insecticides and mosquito repellents) intended to interfere with the transmission of the disease are not highly effective, and the profile of resistance of the parasite to chemoprophylactic and chemotherapeutic agents is increasing. The dawn of the new millennium has seen a resurgence of interest in the disease by government and philanthropic organizations, but we are still faced with compl- ities of the parasite, the host, and the vector, and the interactions among them. Malaria Methods and Protocols offers a comprehensive collection of protocols describing conventional and state-of-the-art techniques for the study of malaria, as well as associated theory and potential problems, written by experts in the field. The major themes reflected here include assessing the risk of infection and severity of disease, laboratory models, diagnosis and typing, molecular biology techniques, immunological techniques, cell biology techniques, and field applications.

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Contents

J Kevin Baird Michael J Bangs Jason D Maguire
13
Maintenance of the Plasmodium berghei Life Cycle
25
Mouse Models for ErythrocyticStage Malaria
57
II Infection of Saimiri and Aotus Monkeys
85
Vector Analysis
95
Nested
103
PCRRFLP Analysis
117
Microsatellite Analysis in Plasmodium falciparum
131
Peptide Vaccination
335
DNA Vaccination
347
Assessing AntigenSpecific CD8+ and CD4+TCell Responses in Mice
361
Assessing CD4+ Helper TLymphocyte Responses by Lymphoproliferation
369
Limiting Dilution Analysis of AntigenSpecific CD4+ TCell Responses
385
Malaria BloodStage ParasiteSpecific CD4+T Cells After
401
Using Flow Cytometry
409
Aftab A Ansari and Ann E Mayne
423

Quantitation of LiverStage Parasites
137
Extraction and Purification of Plasmodium Total
151
Extraction and Purification of Plasmodium Parasite
159
SDSPAGE and Western Blotting of Plasmodium falciparum Proteins
177
Nested PCR Analysis of Plasmodium Parasites
189
RFLP Analysis
205
Analysis of Gene Expression by RTPCR
213
In Situ Detection of RNA in Blood and MosquitoStage Malaria Parasites
225
Daniel J Carucci Paul Horrocks and Malcolm J Gardner 23 Construction of Genomic Libraries from the DNA of Plasmodium Species
235
Cummings Dharmendar Rathore
241
NucleaseTreated Genomic
253
David A Fidock Dharmendar Rathore and Thomas F McCutchan
265
Production of StageSpecific Plasmodium falciparum cDNA Libraries
277
Construction and Screening of YAC Libraries
291
Episomal Transformation of Plasmodium berghei
305
Vandana Thathy and Robert Ménard
317
and José Alejandro López 39 Assessing AntigenSpecific CD8+ CTL Responses in Humans
437
Doolan
445
Human Antibody Subclass ELISA
457
II Analysis
469
In Vitro Culture of Plasmodium Parasites
477
J David Haynes and J Kathleen Moch 45 Hepatic Portal Branch Inoculation
489
John B Sacci Jr
499
Hepatocyte Perfusion Isolation and Culture
503
Inhibition of LiverStage Development Assay
517
Erythrocytic Malaria Growth or Invasion Inhibition Assays with Emphasis
535
Analysis of CSABinding Parasites and Antiadhesion Antibodies
555
Analysis of the Adhesive Properties of Plasmodium falciparumInfected
561
Preparation of Adhesive Targets for FlowBased Cytoadhesion Assays
571
Triton X114 Phase Partitioning for Antigen Characterization
581
Field Trials
607
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