An enzyme-linked immunosorbent assay using monoclonal antibodies for the detection of respiratory syncytial virus in clinical specimens

Abstract

An enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of respiratory syncytial virus in nasopharyngeal secretions. This assay employed as immunoreagents two monoclonal antibodies directed against two distinct epitopes of the viral nucleocapsid. One of them (RSV 4) was used for antigen capture and the other (NC 4) was labelled with N-hydroxy-succinimide-epsilon-caproil biotin and used for antigen detection. Streptavidin biotin-peroxidase complexes were employed as amplification mode. The immunoassay was performed in 6 hours and was able to detect as little as 1 ng/ml of purified nucleocapsid. When 87 nasopharyngeal secretions were analyzed by an indirect immunofluorescence assay using commercial reagents and by the newly developed ELISA, the sensitivity and the specificity of the two assays were found to be very similar.

Incidence of acute otitis media associated with group A and B respiratory syncytial virus infections

“…MAbs RSV4 and NC4 were obtained from G Obert, Inserm, Strasbourg, France. They were generated to RSV strain A2 and recognize the N protein of both RSV groups (19). RS102 was generated to Randall strain of RSV and is specific for the N protein of group A strains only (16).…”

Time-resolved fluoroimmunoassay compared with virus isolation for rapid detection of respiratory syncytial virus in nasopharyngeal aspirates

“…3found a combination of polyclonal serum as a capture antibody and MAbs as detectors superior to MAbs alone in direct-enzyme amplified assays. Only recently, comparable results have been reported by using MAbs both on the soiid phase and as the biotinylated indicator antibody (17).…”

“…MAbs. Hybridomas producing MAbs (RSV4 and NC4) directed against different antigenic sites of the RSV nucleoprotein were developed as described elsewhere (17). Ascites fluid was generated by injecting hybridoma cells intraperitoneally in pristane-primed BALB/c mice.…”

Comparison of monoclonal biotin-avidin enzyme immunoassay and monoclonal time-resolved fluoroimmunoassay in detection of respiratory virus antigens

“…Acute respiratory viral infections are one of the most common causes of seasonal morbidity and of massive absence from work, with resulting economic repercussions. Many groups have reported a number of methods for the rapid diagnosis of such viral respiratory infections (Grandien et al, 1985) as respiratory syncytial virus (Clayton et al, 1987;Obert and Beyer, 1988;Bruckovfi et al, 1989), adenovirus (Lehtom~iki et al, 1986;Bruckovfi et al, 1989), influenzavirus (Bucher et al, 1987;Coonrod et al, 1988;Havlickova et al, 1990), and rhinovirus (A1-Nakib et al, 1989) infections. Moreover, commercially available kits for the detection of respiratory syncytial virus have been evaluated (Freymuth et al, 1986(Freymuth et al, , 1991Halstead et al, 1990).…”