ABSTRACT
This report compares two assay systems for monitoring human immunodeficiency virus (HIV) replication in peripheral blood leukocyte cultures. A commercial enzyme-linked immunoassay detected core antigen (p24) in 80% of cell cultures from HIV-seropositive individuals, whereas 67% of the cell cultures produced detectable levels of reverse transcriptase activity. There were clearly three patterns of reverse transcriptase activity produced, two of which may evade detection without regular sampling and maintaining cell cultures for more than 4 weeks. Once established, core antigen levels remained high so that cell cultures could be confidently monitored by an intermittent screening regimen.
Subject(s)
HIV Antigens/analysis , HIV-1/physiology , Leukocytes/microbiology , RNA-Directed DNA Polymerase/analysis , Retroviridae Proteins/analysis , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , HIV Core Protein p24 , HIV-1/enzymology , HIV-1/immunology , Humans , Predictive Value of Tests , Reagent Kits, Diagnostic , Virus ReplicationABSTRACT
Five commercial assays for detecting HIV antigen were evaluated using a panel of 40 coded samples in six laboratories. All assays were capable of detecting HIV-1 antigen (HIV-1 Ag), and are likely to prove useful for monitoring supernatant fluids from a variety of cell cultures. The concentration of HIV-1 protein which the assays detected varied from 25 ng/ml down to 25 pg/ml. Three of the five assays were also able to detect HIV-2 Ag. More extensive evaluations are needed to determine the sensitivity and specificity of these assays on serum samples and body fluids.
Subject(s)
Antigens, Viral/analysis , HIV/isolation & purification , Viral Proteins/analysis , Antibodies, Viral/immunology , Cells, Cultured , HIV/immunology , HIV Antibodies , Humans , Immunoenzyme Techniques , Lymphocytes/microbiology , Radioimmunoassay/methodsABSTRACT
An enzyme immunoassay (EIA) was developed for detection of human immunodeficiency virus antigen (HIV Ag) in tissue culture supernatants. The assay was found to be specific for HIV and cheaper, easier to perform and more sensitive than the generally used reverse transcriptase (RT) assay. Cultures of peripheral blood leucocytes (PBL) from 106 patients with acquired immune deficiency syndrome (AIDS), AIDS related complex (ARC), healthy anti-HIV positive subjects and healthy anti-HIV negative subjects were held for 35 days and the supernatant fluid tested at regular intervals by EIA and RT. Of these 106 cultures, the presence of HIV was detected by EIA in 27 and by RT in 21. While six cultures were positive by EIA alone, none were positive by RT alone; the specificity of the results in the six EIA positive RT negative cultures was confirmed by subculture. In the 21 cultures in which HIV was detected by both techniques, the EIA became positive first on 10 occasions; in the remaining cultures both tests became positive at the same time. The HIV Ag assay reduces the time taken to process specimens and thus increases the efficiency and reduces the cost of isolation procedures.
