Human Retrovirus Protocols: Virology and Molecular Biology

ISBN-13: 9781588294951 / Angielski / Twarda / 2005 / 494 str.

Tuofu Zhu; Tuofu Zhu

Opis

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.

Recenzje

"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

Spis treści

Part I. 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 Janet D. Siliciano and Robert F. Siliciano Isolation, Propagation, and Titration of Human Immunodeficiency Virus Type 1 From Peripheral Blood of Infected Individuals Hanneke Schuitemaker and Neeltje A. Kootstra Isolation of Human Immunodeficiency Virus Type 1 From Peripheral Blood Monocytes Paul R. Gorry, Secondo Sonza, Katherine Kedzierska, and Suzanne M. Crowe Isolation, Propagation, and HIV-1 Infection of Monocyte-Derived Macrophages and Recovery of Virus From Brain and Cerebrospinal Fluid Santhi Gorantla, Myhanh Che, and Howard E. Gendelman Isolation and HIV-1 Infection of Primary Human Microglia From Fetal and Adult Tissue Kathleen Borgmann, Howard E. Gendelman, and Anuja Ghorpade Isolation of Human Immunodeficiency Virus Type 1 From Semen and Vaginal Fluids Ann A. Kiessling Isolation and Quantification of HIV From Lymph Nodes Catherine Tamalet Isolation of Human Immunodeficiency Virus Type 2 Biological Clones From Peripheral Blood Lymphocytes Hetty Blaak Isolation and Confirmation of Human T-Cell Leukemia Virus Type 2 From Peripheral Blood Mononuclear Cells Michael D. Lairmore and Andy Montgomery Isolation of Foamy Viruses From Peripheral Blood Lymphocytes Joëlle Tobaly-Tapiero, Patricia Bittoun, and Ali Saïb Alu-LTR Real-Time Nested PCR Assay for Quantifying Integrated HIV-1 DNA Audrey Brussel, Olivier Delelis, and Pierre Sonigo Quantification of HFV-Integrated DNA in Human Cells by Alu-LTR Real-Time PCR Olivier Delelis, Audrey Brussel, and Pierre Sonigo Detection of HIV-1 Provirus and RNA by In Situ Amplification Alcina Nicol and Gerard J.Nuovo Detection of HTLV-1 Gene on Cytologic Smear Slides Kenji Kashima, Tsutomu Daa, and Shigeo Yokoyama Detection of HIV-2 by PCR Feng Gao Quantitation of HIV-1 Viral RNA in Blood Plasma and Genital Secretions Susan A. Fiscus Quantification of Proviral DNA Load of Human Immunodeficiency Virus Type 2 Subtypes A and B Using Real-Time PCR Marie Gueudin, Florence Damond, and François Simon Plasma RNA Viral Load in HIV-1 Group O Infection by Real-Time PCR Marie Gueudin and François Simon A New Combined HIV p24 Antigen and Anti-HIV-1/2/O Screening Assay Susanne Polywka, Hedwig Duttmann, Frank Lübben, Rainer Laufs, and Jürgen Feldner A New Automated Fourth-Generation HIV Screening Assay With Sensitive Antigen Detection Module and High Specificity Bernard Weber Detection of Antiviral Antibodies Using Enzyme-Linked Immunosorbent Assay Ingrid G. Winkler Plaque-Reduction Assays for Human and Simian Immunodeficiency Virus Neutralization Anna Nordqvist and Eva Maria Fenyö Detection of Drug-Resistant HIV-1 Strains Ann A. Kiessling, S. J. Eyre, and B. D. Desmarais Part II. Molecular Biology Methods Determination of Cell Tropism of HIV-1 Neeltje A. Kootstra and Hanneke Schuitemaker Determination of Co-Receptor Usage of HIV-1 Hanneke Schuitemaker and Neeltje A. Kootstra Quantitative Evaluation of HIV and SIV Co-Receptor Use With GHOST(3) Cell Assay Dalma Vödrös and Eva Maria Fenyö Phenotypic Characterization of Blood Monocytes From HIV-Infected Individuals Philip J. Ellery and Suzanne M. Crowe Methods to Determine HIV-1 Ex Vivo Fitness Awet Abraha, Ryan M. Troyer, Miguel E. Quiñones-Mateu, and Eric J. Arts A Yeast Recombination-Based Cloning System to Produce Chimeric HIV-1 Viruses and Express HIV-1 Genes Dawn

Blurb

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™ 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.