Development of a Monoclonal Antibody-Based p24 Capsid Antigen Detection Assay for HTLV-I, HTLV-II, and STLV-I Infection
- 1 April 1992
- journal article
- research article
- Published by Mary Ann Liebert Inc in AIDS Research and Human Retroviruses
- Vol. 8 (4), 527-532
- https://doi.org/10.1089/aid.1992.8.527
Abstract
A monoclonal antibody-based antigen capture enzyme-linked immunosorbent assay (ELISA) was developed and employed to detect p24 capsid antigen from human T-cell lymphotropic viruses type I and II (HTLV-I, HTLV-II), simian T-cell lymphotropic virus type I (STLV-I) -infected cell lines, and from mononuclear cell cocultures of HTLV-infected humans and STLV-I infected monkeys. A monoclonal antibody specific for HTLV p24 and p53 capsid antigens was coated onto 96-well microtiter plates to capture HTLV/STLV antigen. Captured antigen was then detected by the addition of a polyclonal, biotinylated human anti-HTLV-I antibody, and color developed with tetramethyl benzidine/H202 substrate. As little as 15 pg/ml of HTLV-I p24 antigen could be detected in this assay. Culture supernatants from HTLV-I-infected cell lines (HUT-102, MT-2, C5/MJ), HTLV-II-infected cell lines (Mo-T, Mo-B, PanG 12.1, NRA) and STLV-I-infected cell lines (Matsu, NEPC M39) were all positive in the assay. In addition, p24 was detected from peripheral blood mononuclear cell (PBMC) cocultures of 8 of 8 (100%) HTLV-I diseased patients, 14 of 20 (70%) HTLV-I and HTLV-II-infected, asymptomatic persons, and 8 of 8 (100%) STLV-I-infected, asymptomatic monkeys. Culture supernatants of cells infected with human immunodeficiency virus type (HIV-1), simian immunodeficiency virus (SIV), Chlamydia trachomatis, cytomegalovirus (CMV), herpes simplex I and II (HSV), feline leukemia virus (FELV), bovine leukemia virus (BLV), and bovine immunodeficiency virus (BIV) were all negative. Similarly, normal human peripheral blood mononuclear cells and uninfected, transformed human T cells, were also negative in the assay. The antigen assay is a sensitive and specific method for the detection of HTLV-I, HTLV-II, and STLV-I p24 capsid antigen and can replace reverse transcriptase assays in the confirmation of tissue cultures for these retroviruses.Keywords
This publication has 27 references indexed in Scilit:
- Relative prevalence and risk factors of HTLV-I and HTLV-II infection in US blood donorsThe Lancet, 1991
- HTLV-I, Adult T-Cell leukemia, and tropical spastic paraparesisAnnals of Neurology, 1988
- A Second Isolate of HTLV-II Associated with Atypical Hairy-Cell LeukemiaNew England Journal of Medicine, 1986
- Isolation of T-Cell Tropic HTLV-III-Like Retrovirus from MacaquesScience, 1985
- Isolation and Transmission of Human Retrovirus (Human T-Cell Leukemia Virus)Science, 1983
- Isolation and characterization of retrovirus from cell lines of human adult T-cell leukemia and its implication in the disease.Proceedings of the National Academy of Sciences, 1982
- Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphomaProceedings of the National Academy of Sciences, 1980
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- Terminal deoxynucleotidyl transferase activity in a cell line (Molt-4) derived from the peripheral blood of a patient with acute lymphoblastic leukemiaBiochemical and Biophysical Research Communications, 1973
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970