Humana Human Retrovirus Protocols: Virology and Molecular Biology: 304 (Methods in Molecular Biology, 304)

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 "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 From the 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™ 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 an