A comprehensive compilation of research techniques necessary for investigating the virology, immunology and molecular biology of HIV-1. Protocols are also provided which represent state of the art approaches to a wide spectrum of HIV related issues.

Inhalt

Part 1 Antibody detection in patient's serum: ELISA assay; western blot analysis. Part 2 Virus detection in pathological specimen: p24 assay; polymerase chain reaction (PCR); in situ hybridization; southern blot analysis. Part 3 Virus isolation and production: isolation from periperal blood; isolation from tissues; virus production in stable cell lines. Part 4 Virus titration: on MT4 cells; on HeLa cells; on C81-66; on SUP T1. Part 5 Virus detection in cell lines: reverse transcriptase assay; p24 assay; immunofluorescence (IF); cythopathic effect (CPE); radioimmunoprecipitation (RIP). Part 6 Virus purification and storage. Part 7 Proviral DNA cloning: recombinant DNA libraries; polymerase chain reaction (PCR). Part 8 Tranfection of biologically active molecular clones: DEAE dextran; electroporation; CaPO4 transfection; lipofection; selection of stably transfected cell lines. Part 9 Host immune response: neutralizing antibodies; ADCC/CMC; cytotoxity - CTL; proliferative response. Part 10 Cytokine assays. Part 11 Antiviral drugs assays. Appendices: source of reagents; biosafey issues.