HIV Protocols

1. Front Matter

   Pages i-xiii

   PDF

2. Virology

   1. Isolation and Expansion of HIV from Cells and Body Fluids by Coculture

      • James R. Lane

      Pages 3-10

   2. Quantitative HIV Culture

      • James R. Lane

      Pages 11-15

   3. Quantitation of Cell-Free HIV by Reverse Transcriptase Activity

      • James R. Lane

      Pages 17-22

   4. Determination of Syncytium-Inducing Phenotype of Primary HIV-1 Isolates Using MT-2 cells

      • Mark K. Louder, John R. Mascola

      Pages 23-27

   5. Determination of HIV-1 Chemokine Coreceptor Tropism Using Transduced Human Osteosarcoma (HOS) Cells

      • Mark K. Louder, John R. Mascola

      Pages 29-33

   6. Use of Luciferase Reporter Viruses for Studying HIV Entry

      • Rong Liu, Benjamin Chen, Nathaniel R. Landau

      Pages 35-40

   7. A Cell-Cell Fusion Assay to Monitor HIV-1 Env Interactions with Chemokine Receptors

      • Aimee L. Edinger, Robert W. Doms

      Pages 41-49

   8. Methods of Culturing HIV-1 from Semen

      • Bruce L. Gilliam

      Pages 51-57

3. Molecular Biology

   1. Detection of HIV-1 Nucleic Acids by Southern Blotting

      • Richard A. McDonald

      Pages 61-69

   2. Detection of HIV-1 Nucleic Acids by Northern Blotting

      • Richard A. McDonald, Christopher A. D. Smith

      Pages 71-81

   3. Rapid Identification of Cloned HIV-1 Fragments

      • Wei Dong, Anindita Kar Roy, Yen Li

      Pages 83-87

   4. PCR Amplification and Cloning of Virtually Full-Length HIV-1 Provirus

      • Mika Salminen

      Pages 89-98

   5. Quantitation of HIV-1gag DNA and RNA From Single Frozen Cell Pellets

      • Maryanne Vahey, Sandra C. Barrick, Martin Nau

      Pages 99-118

   6. Quantitative PCR for HIV-1 Proviral DNA

      • Davide Zella, Paola Secchiero

      Pages 119-124

   7. Diagnosis and Direct Automated Sequencing of HIV-1 From Dried Blood Spots (DBS) Collected on Filter Paper

      • Sharon A. Cassol, Stanley Read, Bruce G. Weniger, Richard Pilon, Barbara Leung, Theresa Mo

      Pages 125-138

   8. Quantification of HIV-1 RNA in Dried Plasma Spots (DPS)

      • Sharon A. Cassol, Francisco Diaz-Mitoma, Richard Pilon, Michelle Janes, D. William Cameron

      Pages 139-149

   9. Quantitation of HIV-1 RNA in Genital Secretions

      • Bruce L. Gilliam

      Pages 151-164

   10. Detection of Nucleic Acids in Cells and Tissues by In Situ Polymerase Chain Reaction

       • Omar Bagasra, Lisa E. Bobroski, Mohammad Amjad, Matthew Memoli, Maureen V. Abbey

       Pages 165-183

   11. Determination of the Promoter Activity of HIV-1 Using the Chloramphenicol Acetyltransferase Reporter Gene Assay

       • Loretta Tse, Nelson L. Michael

       Pages 185-196

The worldwide impact of infection with human immunodeficiency virus type 1 (HIV- is reflected in the cumulative number ofHIV- 1 infections, which is now predicted to exceed 40 million by the year 2000---equivalent to the n- ber of humans who perished in World War II. The medical and scientific - sponse to the HIV-1 pandemic has steadily grown since its recognition in 1981. The outlay by the United States alone for HIV research funded by the National Institutes of Health in 1997 was $1. 4 billion. Laboratory-based HIV research has brought together academic clinicians, retrovirologists, molecular biologists, and immunologists in the formation of research teams attempting to dissect the viral and host factors contributing to disease pathogenesis. Increasing focus is being placed on those aspects of viral biology and host immune responses that bear on the development of vaccines to prevent HIV infection. HIVProtocols reflects the state of HIV research in several ways. First, chapters are organized into four sections: Virology, Molecular Biology, Humoral Immunology, and Cellular Immunology. This organization is a natural consequence of the diverse scientific disciplines that have been attracted to HIV research. Second, the chapters reflect such diverse research directions as viral coreceptor usage, quantitation of viral genomes, HIV promoter function, B-cell epitope mapping, and measurements of T-cell function, each of which bears on the goal of understanding the viral and host immune responses that will be critical to the design of effective preventive vaccines.

". . . a great companion to the more general laboratory protocol manuals. . . provide[s] an impressively up-to-date collection of laboratory methods for every common and not-so-common AIDS lab experiment."-Antiviral Research

HIV Protocols is a good addition to a basic science laboratory that processes various HIV clinical specimens. Graduate students or clinical fellows starting out on their careers in basic experimental science will find it most useful....Overall, this book compiles a number of general basic science protocols in the fields of virology, molecular biology and immunology as applied to the study of HIV. The thorough nature of the materials and methods section of each chapter ensures that the reader will get off to a good start when using these techniques in their own laboratory." -Can J Infect Dis

• Walter Reed Army Institute of Research, Rockville

  Nelson L. Michael

• Henry M. Jackson Foundation, Rockville

  Jerome H. Kim

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