Human Retrovirus Protocols: Virology and Molecular Biology by Tuofu Zhu (English

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There are also assays for determining the cell tropism of HIV-1, the coreceptor usage of HIV-1, and human gene expression with HIV-1 infection by microarrays, as well as for phenotyping HIV-1 infected monocytes and examining their fitness.

FORMATHardcover LANGUAGEEnglish CONDITIONBrand New Publisher Description

This volume focuses on the methodologies of virology and moleculr biology of human retrovirus. Organized in two sections, the first deals primarily with methods for the isolation and detection of human retroviruses while the second focuses primarily on the phenotype of human retroviruses. All chapters offer in detail the individual steps necessary to carry out the assays, and provide discussion of problems and pitfalls that may be encountered.

Back Cover

Advances in our understanding of human retroviruses have been occurring at a very rapid pace and have contributed significantly to the development of new biotechnologies. In Human Retrovirus Protocols: Virology and Molecular Biology, recognized international researchers describe in detail the basic and state-of-the-art methods they have optimized for investigating the molecular biology of this class of retrovirus. These readily reproducible techniques range from proven methods for the isolation and detection of human retroviruses to advanced methods for exploring the interplay between the viruses and the host. Here, the researcher will find up-to-date techniques for the isolation and propagation of HIV, HTLV, and foamy virus from peripheral blood mononuclear cells (PBMC), blood monocytes, brain tissues, cerebrospinal fluids, semen, the vagina, and lymph nodes. There are also assays for determining the cell tropism of HIV-1, the coreceptor usage of HIV-1, and human gene expression with HIV-1 infection by microarrays, as well as for phenotyping HIV-1 infected monocytes and examining their fitness. Highlights include the detection and quantification of HIV-1 in resting CD4+, a new cloning system for making recombinant virus, cDNA microarrays, and the determination of genetic polymorphisms in two recently identified HIV-1 co-factors that are critical for HIV-1 infection. The protocols follow the highly praised Methods in Molecular Biology(tm) series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. State-of-the-art and highly practical, Human Retrovirus Protocols: Virology and Molecular Biology offers novice and experienced researchers alike proven tools essential for productive study and understanding of the human retroviruses.

Table of Contents

Methods for Virus Isolation and Detection.- Enhanced Culture Assay for Detection and Quantitation of Latently Infected, Resting CD4+ T-Cells Carrying Replication-Competent Virus in HIV-1-Infected Individuals.- Isolation, Propagation, and Titration of Human Immunodeficiency Virus Type 1 From Peripheral Blood of Infected Individuals.- Isolation of Human Immunodeficiency Virus Type 1 From Peripheral Blood Monocytes.- Isolation, Propagation, and HIV-1 Infection of Monocyte-Derived Macrophages and Recovery of Virus From Brain and Cerebrospinal Fluid.- Isolation and HIV-1 Infection of Primary Human Microglia From Fetal and Adult Tissue.- Isolation of Human Immunodeficiency Virus Type 1 From Semen and Vaginal Fluids.- Isolation and Quantification of HIV From Lymph Nodes.- Isolation of Human Immunodeficiency Virus Type 2 Biological Clones From Peripheral Blood Lymphocytes.- Isolation and Confirmation of Human T-Cell Leukemia Virus Type 2 From Peripheral Blood Mononuclear Cells.- Isolation of Foamy Viruses From Peripheral Blood Lymphocytes.- Alu-LTR Real-Time Nested PCR Assay for Quantifying Integrated HIV-1 DNA.- Quantification of HFV-Integrated DNA in Human Cells by Alu-LTR Real-Time PCR.- Detection of HIV-1 Provirus and RNA by In Situ Amplification.- Detection of HTLV-1 Gene on Cytologic Smear Slides.- Detection of HIV-2 by PCR.- Quantitation of HIV-1 Viral RNA in Blood Plasma and Genital Secretions.- Quantification of Proviral DNA Load of Human Immunodeficiency Virus Type 2 Subtypes A and B Using Real-Time PCR.- Plasma RNA Viral Load in HIV-1 Group O Infection by Real-Time PCR.- A New Combined HIV p24 Antigen and Anti-HIV-1/2/O Screening Assay.- A New Automated Fourth-Generation HIV Screening Assay With Sensitive Antigen Detection Module and High Specificity.- Detection of Antiviral Antibodies Using Enzyme-Linked Immunosorbent Assay.- Plaque-Reduction Assays for Human and Simian Immunodeficiency Virus Neutralization.- Detection of Drug-Resistant HIV-1 Strains.- Molecular Biology Methods.- Determination of Cell Tropism of HIV-1.- Determination of Co-Receptor Usage of HIV-1.- Quantitative Evaluation of HIV and SIV Co-Receptor Use With GHOST(3) Cell Assay.- Phenotypic Characterization of Blood Monocytes From HIV-Infected Individuals.- Methods to Determine HIV-1 Ex Vivo Fitness.- A Yeast Recombination-Based Cloning System to Produce Chimeric HIV-1 Viruses and Express HIV-1 Genes.- PCR Amplification, Cloning, and Construction of HIV-1 Infectious Molecular Clones From Virtually Full-Length HIV-1 Genomes.- Amplification and Cloning of Near Full-Length HIV-2 Genomes.- Growth and Manipulation of a Human T-Cell Leukemia Virus Type 2 Full-Length Molecular Clone.- Construction and Analysis of Genomic, Full-Length Infectious Foamy Virus DNA Clones.- Molecular Characterization of Proteolytic Processing of the Gag Proteins of Human Spumaretrovirus.- Assessing the Relative Efficacy of Antiretroviral Activity of Different Drugs on Macrophages.- Gene Expression Profiling of HIV-1 Infection Using cDNA Microarrays.- Determination of DC-SIGN and DC-SIGNR Repeat Region Variations.

Review

"More than 20 years after the discoveries of HTLV and HIV, there appears to be a need for a consolidated reference of many of the standard and not-so-standard protocols used to characterize the human retroviruses. This book, containing some 37 detailed protocols provided by multinational authors in two parts, does an admirable job. Part I, therefore, includes guidance on isolation of HTLV, HIV and Human Foamy Virus (HFV) from peripheral blood, lymph nodes, CNS tissue and genital secretions (semen, vaginal fluids). It also describes a variety of molecular methods (largely PCR-based) to assess viral levels, integration status and drug sensitivity, plus quantification by antigen detection. At some 313 pages, Part I forms the substantial portion of the book. Part II (168 pages) guides on assessing cell and receptor tropism, construction and utilization of molecular clones and also use of microarrays to look at host and viral gene expression. The book on the whole is well illustrated with clear photographs and diagrams to support the detailed methodology, and the references at the end of each protocol are comprehensive. "-SGM Quarterly

Long Description

A cutting-edge collection of basic and state-of-the-art methods optimized for investigating the molecular biology of this class of retrovirus. These readily reproducible techniques range from methods for the isolation and detection of human retroviruses to cutting-edge methods for exploring the interplay between the viruses and the host. Here, the researcher will find up-to-date techniques for the isolation and propagation of HIV, HTLV, and foamy virus from a variety of sources. There are also assays for determining the cell tropism of HIV-1, the coreceptor usage of HIV-1, and human gene expression with HIV-1 infection by microarrays, as well as for phenotyping HIV-1 infected monocytes and examining their fitness. Highlights include the detection and quantification of HIV-1 in resting CD4+, a new cloning system for making recombinent virus, cDNA microarrays, and the determination of genetic polymorphisms in two recently identified HIV-1 co-factors that are critical for HIV-1 infection.

Review Text

"More than 20 years after the discoveries of HTLV and HIV, there appears to be a need for a consolidated reference of many of the standard and not-so-standard protocols used to characterize the human retroviruses. This book, containing some 37 detailed protocols provided by multinational authors in two parts, does an admirable job. Part I, therefore, includes guidance on isolation of HTLV, HIV and Human Foamy Virus (HFV) from peripheral blood, lymph nodes, CNS tissue and genital secretions (semen, vaginal fluids). It also describes a variety of molecular methods (largely PCR-based) to assess viral levels, integration status and drug sensitivity, plus quantification by antigen detection. At some 313 pages, Part I forms the substantial portion of the book. Part II (168 pages) guides on assessing cell and receptor tropism, construction and utilization of molecular clones and also use of microarrays to look at host and viral gene expression. The book on the whole is well illustrated with clear photographs and diagrams to support the detailed methodology, and the references at the end of each protocol are comprehensive. "-SGM Quarterly

Review Quote

"More than 20 years after the discoveries of HTLV and HIV, there appears to be a need for a consolidated reference of many of the standard and not-so-standard protocols used to characterize the human retroviruses. This book, containing some 37 detailed protocols provided by multinational authors in two parts, does an admirable job. Part I, therefore, includes guidance on isolation of HTLV, HIV and Human Foamy Virus (HFV) from peripheral blood, lymph nodes, CNS tissue and genital secretions (semen, vaginal fluids). It also describes a variety of molecular methods (largely PCR-based) to assess viral levels, integration status and drug sensitivity, plus quantification by antigen detection. At some 313 pages, Part I forms the substantial portion of the book. Part II (168 pages) guides on assessing cell and receptor tropism, construction and utilization of molecular clones and also use of microarrays to look at host and viral gene expression. The book on the whole is well illustrated with clear photographs and diagrams to support the detailed methodology, and the references at the end of each protocol are comprehensive. "-SGM Quarterly

Feature

Includes supplementary material: sn.pub/extras

Details ISBN1588294951 Author Tuofu Zhu Language English ISBN-10 1588294951 ISBN-13 9781588294951 Media Book Format Hardcover Series Number 304 Year 2005 Subtitle Virology and Molecular Biology Place of Publication Totowa, NJ Country of Publication United States Imprint Humana Press Inc. Short Title HUMAN RETROVIRUS PROTOCOLS 200 Pages 494 DOI 10.1604/9781588294951;10.1385/1592599079;10.1007/978-1-59259-907-3 AU Release Date 2005-05-17 NZ Release Date 2005-05-17 US Release Date 2005-05-17 UK Release Date 2005-05-17 Edited by Tuofu Zhu Publisher Humana Press Inc. Edition Description 2005 ed. Series Methods in Molecular Biology Edition 2005th Publication Date 2005-05-17 Alternative 9781617375996 DEWEY 616.0194 Illustrations XIV, 494 p. Audience Professional & Vocational We've got this

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