Rapid viral diagnosis of acute respiratory infections: comparison of enzyme-linked immunosorbent assay and the immunofluorescence technique for detection of viral antigens in nasopharyngeal secretions

Abstract

Nasopharyngeal secretions from adults and children were obtained in Stockholm, Sweden, for routine diagnosis of influenza A virus, influenza B virus, respiratory syncytial (RS) virus, parainfluenza type 3 virus, and adenovirus infections by demonstration of viral antigens directly in the specimens. The cells in nasopharyngeal secretions were pelleted by centrifugation for preparation of cell deposits for diagnosis by the immunofluorescence technique (IF) in London, England, and in Stockholm, whereas the supernatants were used to diagnose infection by the enzyme-linked immunosorbent assay (ELISA) in Stockholm. Titrations of the various purified viruses showed that ELISA could detect viral antigens in amounts corresponding to 1 to 10 ng of virus protein per test well. In a series of 73 specimens tested for influenza A, RS, and parainfluenza type 3 viruses by IF in London and by ELISA in Stockholm, 15 of 18 RS, 14 of 15 influenza A, and 2 of 2 parainfluenza type 3 viral infections were diagnosed by ELISA as compared with IF, giving sensitivities for RS and influenza A viral diagnosis of 83 and 93%, respectively, and a specificity of 100%. In another series of specimens from 35 patients tested for influenza B virus and adenovirus, five influenza B virus and four adenovirus infections were diagnosed by both methods; one additional influenza B infection was detected only by IF and another only by ELISA. Comparisons of diagnostic results between the two methods performed in Stockholm gave nonagreement of results for 37 of 1,593 tests (2.5%) for the five viruses. The conclusion reached was that the described ELISA, although a satisfactory test, had somewhat less sensitivity than did IF for the detection of respiratory viral infections. This could possibly be explained by unnecessary dilutions of specimens at the time of collection; transportation, processing, and storage of specimens were less complicated than for IF.

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

“…What is the advantage of Biotin-EIA or TR-FIA compared with the immunofluorescence technique (Gardner and McQuillin, 1980;Grandien et al, 1985) in rapid diagnosis of respiratory infections? The answer depends on the location of the laboratory, the number of specimens tested daily, the availability of reagents, and the previous expertise of the laboratory.…”

“…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).…”

Antigen detection in human respiratory Coronavirus infections by monoclonal time-resolved fluoroimmunoassay

“…Fluorescent antibody tests -the first rapid antigen tests developed for respiratory viruses -are still widely used in clinical laboratories and have been successfully applied to the human respiratory coronaviruses as well (McIntosh et al, 1978). Enzyme immunoassays have become even more important for rapid viral diagnosis, because EIA is more sensitive than IFA and is therefore the critical test with which to compare any new test (Halonen et al, 1983(Halonen et al, , 1985Grandien et al, 1985;Hornsleth et al, 1986;Hughes et al, 1988;Hierholzer et al, 1990;Takimoto et al, 1991;McIntosh et al, 1993). Thus, this study included optimized EIA tests with monoclonal and polyclonal antibodies in a direct comparison with optimized TR-FIA tests with monoclonal antibodies.…”

La grippe saisonnière

“…Le plus souvent, la technique est la même, et les diffé rences portent sur la pré paration de l'antigè ne et le type d'anticorps. Les antigè nes viraux extraits des pré lè vements sont capté s par un anticorps monoclonal dans les puits d'une plaque de microtitration, et leur dé tection se fait par un test EIA classique [33,36]. Leur sensibilité comparé e à l'isolement en culture varie de 50 à 90 % selon les auteurs [37].…”