p24 antigen capture assay for quantification of human immunodeficiency virus using readily available inexpensive reagents.
Antigen capture enzyme-linked immunosorbent assay (ELISA) for quantitation of the p24 gag protein of human immunodeficiency virus type-1 (HIV-1) is currently the most common method used to demonstrate virus replication both in vivo and in vitro. The present paper describes an ELISA employing readily available inexpensive reagents and gives detailed suggestions for optimizing the variables for specific purposes. The assay is as sensitive as commercial kits (25 pg/ml) and has a linear dose response over a wide range of p24 concentrations (25-1000 pg/ml). For these reasons, as well as its low cost, this assay has proven useful in a variety of research applications. This assay also has been found to be effective in detecting the gag protein of human immunodeficiency virus type-2 and simian immunodeficiency virus.
Wehrly
K
K
Laboratory of Persistent, Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, Montana 59840-2932, USA.
Chesebro
B
B
D001426:Bacterial Proteins / Q000032:analysis; D001560:Benzidines / Q000032:analysis; D001710:Biotin / Q000737:chemistry; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015683:Gene Products, gag / Q000032:analysis; D016655:HIV Core Protein p24 / Q000032:analysis*; D015497:HIV-1 / Q000276:immunology / Q000302:isolation & purification*; D015498:HIV-2 / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016756:Immunoglobulins, Intravenous; D007202:Indicators and Reagents / Q000191:economics; D010544:Peroxidases / Q000032:analysis; D015302:Simian Immunodeficiency Virus / Q000302:isolation & purification; D019809:Streptavidin
D001426:Bacterial Proteins; D001560:Benzidines; D015683:Gene Products, gag; D016655:HIV Core Protein p24; C084715:HIV hyperimmune globulin; D016756:Immunoglobulins, Intravenous; D007202:Indicators and Reagents; C021758:3,3',5,5'-tetramethylbenzidine; D001710:Biotin; D019809:Streptavidin; D010544:Peroxidases