Use of monoclonal antibodies for the detection and quantitation of HIV1 core protein p25: comparative evaluation of in vitro HIV1 infection by immunofluorescence, antigen capture ELISA and reverse transcriptase assays.
A two-site enzyme linked immunosorbent assay (ELISA) was developed to detect and quantify the HIV1 core protein p25 in the cell-free supernatant from virus-infected CEM cell culture, and compared with other assays. The assay, based on a sandwich method, employs two monoclonal antibodies (mAb) directed against different epitopes on the p25 core protein of HIV1, one used for p25 antigen capture and the other as a biotinylated probe. This immunoassay is sensitive enough to detect as little as 30 pg/ml of recombinant p25, the range of sensitivity of commercial kits, and therefore compares favourably with the conventional reverse transcriptase assay. Moreover, several hundred assays can be monitored quite conveniently by this simple ELISA procedure, which represents a useful tool for detection of HIV1 replication in microculture systems and rapid screening of antiretroviral agents using the reference strain HIV1-BRU as a model system.
Tatsumi
M
M
INSERM U322, Unité de Recherches sur les Rétrovirus et Maladies Associées, Marseille, France.
Jean
F
F
Robert
V
V
Chermann
J C
JC
Devaux
C
C
D000163:Acquired Immunodeficiency Syndrome / Q000175:diagnosis*; D000911:Antibodies, Monoclonal / Q000276:immunology; D001710:Biotin; D002478:Cells, Cultured; D000090266:Chromobox Protein Homolog 5; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005455:Fluorescent Antibody Technique; D015683:Gene Products, gag / Q000032:analysis* / Q000276:immunology; D015488:HIV Antigens / Q000276:immunology; D015497:HIV-1 / Q000032:analysis* / Q000254:growth & development; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D012194:RNA-Directed DNA Polymerase; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014779:Virus Replication; D054301:gag Gene Products, Human Immunodeficiency Virus
D000911:Antibodies, Monoclonal; C000715594:CBX5 protein, human; D015683:Gene Products, gag; D015488:HIV Antigens; C059827:HIV gag protein p25; D011933:Reagent Kits, Diagnostic; D054301:gag Gene Products, Human Immunodeficiency Virus; D000090266:Chromobox Protein Homolog 5; D001710:Biotin; D012194:RNA-Directed DNA Polymerase