64] Sensitive immunoassay for human immunodeficiency viral core proteins

Abstract

Publisher Summary This chapter discusses the role of sensitive immunoassay for human immunodeficiency viral core proteins. The development of sensitive assays is a critical need in improving the detection, diagnosis, and treatment of disease. It is particularly important in diseases with devastating ramifications, such as acquired immunodeficiency syndrome (AIDS). The currently used blood-screening tests are capable of detecting antibodies to human immunodeficiency virus (HIV), the causative viral agent of AIDS. Highly specific rabbit polyclonal antisera provide the capture and detector immunoglobin reagents. Inactivated viral lysate serves as the standard. A biotin–streptavidin–horseradish peroxidase (HRP) couple provides the probe in a microtiter plate sandwich enzyme-linked immunoassay (ELISA) format. When the assay is used to detect viral antigen in supernatants of amplified cultures of peripheral blood lymphocytes, direct comparisons with the reverse transcriptase assay have shown an excellent correlation with additional advantages of increased sensitivity (up to 1000-fold) and reproducibility. The specificity of a positive color response generated in the HIV p24 ELISA is confirmed by specific inhibition of signal by preincubating the sample with human antibody to HIV p24.