Malaria Methods and Protocols

Buch | Softcover

631 Seiten

2010 | Softcover reprint of hardcover 1st ed. 2002
Humana Press Inc. (Verlag)
978-1-61737-205-6 (ISBN)

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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.

Assessing Risk of Infection and Severity of Disease.- Vector Incrimination and Entomological Inoculation Rates.- Epidemiological Measures of Risk of Malaria.- Laboratory Models.- Maintenance of the Plasmodium berghei Life Cycle.- Mouse Models for Pre-Erythrocytic-Stage Malaria.- Mouse Models for Erythrocytic-Stage Malaria.- Nonhuman Primate Models.- Nonhuman Primate Models.- Diagnosis and Typing.- Vector Analysis.- Genotyping of Plasmodium spp..- Genotyping of Plasmodium falciparum.- Microsatellite Analysis in Plasmodium falciparum.- Quantitation of Liver-Stage Parasites by Automated TaqMan® Real-Time PCR.- Quantitation of Liver-Stage Parasites by Competitive RT-PCR.- Molecular Biology Techniques.- Extraction and Purification of Plasmodium Total RNA.- Extraction and Purification of Plasmodium Parasite DNA.- Southern Blotting of Parasite DNA.- SDS-PAGE and Western Blotting of Plasmodium falciparum Proteins.- Nested PCR Analysis of Plasmodium Parasites.- RFLP Analysis.- Analysis of Gene Expression by RT-PCR.- In Situ Detection of RNA in Blood- and Mosquito-Stage Malaria Parasites.- Purification of Chromosomes from Plasmodium falciparum.- Construction of Genomic Libraries from the DNA of Plasmodium Species.- Construction of a Gene Library with Mung Bean Nuclease-Treated Genomic DNA.- Construction of Plasmodium falciparum ? cDNA Libraries.- Production of Stage-Specific Plasmodium falciparum cDNA Libraries Using Subtractive Hybridization.- Construction and Screening of YAC Libraries.- Episomal Transformation of Plasmodium berghei.- Gene Targeting in Plasmodium berghei.- Immunological Techniques.- Peptide Vaccination.- DNA Vaccination.- Assessing Antigen-Specific CD8+ and CD4+ T-Cell Responses in Mice After Immunization with Recombinant Viruses.- Assessing CD4+HelperT-Lymphocyte Responses by Lymphoproliferation.- Limiting Dilution Analysis of Antigen-Specific CD4+ T-Cell Responses in Mice.- Cell Trafficking.- Assessing Antigen-Specific Proliferation and Cytokine Responses Using Flow Cytometry.- Cytokine Analysis by Intracellular Staining.- Assessment of Antigen-Specific CTL- and CD8+-Dependent IFN-? Responses in Mice.- Assessing Antigen-Specific CD8+ CTL Responses in Humans.- Human Antibody Subclass ELISA.- Systemic Nitric Oxide Production in Human Malaria.- Systemic Nitric Oxide Production in Human Malaria.- Cell Biology Techniques.- In Vitro Culture of Plasmodium Parasites.- Automated Synchronization of Plasmodium falciparum Parasites by Culture in a Temperature-Cycling Incubator.- Hepatic Portal Branch Inoculation.- Hepatocyte Perfusion, Isolation, and Culture.- Inhibition of Sporozoite Invasion.- Inhibition of Liver-Stage Development Assay.- T-Cell Mediated Inhibition of Liver-Stage Development Assay.- Antibody-Dependent Cellular Inhibition Assay.- Erythrocytic Malaria Growth or Invasion Inhibition Assays with Emphasis on Suspension Culture GIA.- Analysis of CSA-Binding Parasites and Antiadhesion Antibodies.- Analysis of the Adhesive Properties of Plasmodium falciparum-Infected Red Blood Cells Under Conditions of Flow.- Preparation of Adhesive Targets for Flow-Based Cytoadhesion Assays.- Triton X-114 Phase Partitioning for Antigen Characterization.- Immunoprecipitation for Antigen Localization.- Testing Interventions For Malaria Control.- Field Trials.

Reihe/Serie	Methods in Molecular Medicine ; 72
Zusatzinfo	XV, 631 p.
Verlagsort	Totowa, NJ
Sprache	englisch
Themenwelt	Medizin / Pharmazie ► Medizinische Fachgebiete
Themenwelt	Studium ► Querschnittsbereiche ► Infektiologie / Immunologie
ISBN-10	1-61737-205-6 / 1617372056
ISBN-13	978-1-61737-205-6 / 9781617372056
Zustand	Neuware