Abstract
The horseradish peroxidase-catalyzed oxidation of luminol was used in a chemiluminescence avidin-biotin enzyme-linked immunosorbent assay (CL-ABE) to detect respiratory syncytial virus (RSV) and rotavirus (ROV). This CL-ABE was carried out in a new apparatus constructed for the photographic registration of the light emission produced. When measured in a spectrophotometer, the light emission produced by the oxidation of luminol showed a peak emission at 425 nm. The chemiluminescence output reached maximum within 1 min after the initiation of luminol oxidation and diminished to one-half of maximum emission within 8 min. When titrations of purified ROV by avidin-biotin enzyme-linked immunosorbent assay (ABE) were monitored by CL-ABE and by conventional orthophenylenediamine (OPD) staining (OPD-ABE), the detection limits were 0.01 and 0.04 ng of ROV protein, respectively. Similar titrations of purified RSV gave detection limits of 0.2 and 0.8 ng of RSV protein by CL-ABE and OPD-ABE, respectively. When 26 RSV-positive samples of nasopharyngeal secretions (NPS) were titrated by CL-ABE and by OPD-ABE, the mean titers obtained were 737 and 254, respectively. When 19 ROV-positive fecal samples were titrated by CL-ABE and by OPD-ABE, the mean titers obtained were 82,000 and 29,000, respectively. When samples of NPS from 123 infants and children with acute lower respiratory disease were tested for the presence of RSV, 31 NPS samples were RSV positive by CL-ABE, and 29 of these 31 NPS samples were RSV positive by OPD-ABE.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Bunce R. A., Thorpe G. H., Gibbons J. E., Killeen P. R., Ogden G., Kricka L. J., Whitehead T. P. Camera luminometer for use with luminescent assays. Analyst. 1985 Jun;110(6):657–663. doi: 10.1039/an9851000657. [DOI] [PubMed] [Google Scholar]
- Cormier M. J., Prichard P. M. An investigation of the mechanifm of the luminescent peroxidation of luminol by stopped flow techniques. J Biol Chem. 1968 Sep 25;243(18):4706–4714. [PubMed] [Google Scholar]
- Grauballe P. C., Vestergaard B. F., Meyling A., Genner J. Optimized enzyme-linked immunosorbent assay for detection of human and bovine rotavirus in stools: Comparison with electron-microscopy, immunoelectro-osmophoresis, and fluorescent antibody techniques. J Med Virol. 1981;7(1):29–40. doi: 10.1002/jmv.1890070104. [DOI] [PubMed] [Google Scholar]
- Hornsleth A., Friis B., Andersen P., Brenøe E. Detection of respiratory syncytial virus in nasopharyngeal secretions by ELISA: comparison with fluorescent antibody technique. J Med Virol. 1982;10(4):273–281. doi: 10.1002/jmv.1890100407. [DOI] [PubMed] [Google Scholar]
- Hornsleth A., Friis B., Krasilnikof P. A. Detection of respiratory syncytial virus in nasopharyngeal secretions by a biotin-avidin ELISA more sensitive than the fluorescent antibody technique. J Med Virol. 1986 Feb;18(2):113–117. doi: 10.1002/jmv.1890180203. [DOI] [PubMed] [Google Scholar]
- Hornsleth A., Grauballe P. C., Friis B., Genner J., Pedersen I. R. Production of antiserum to respiratory syncytial virus polypeptides: application in enzyme-linked immunosorbent assay. J Clin Microbiol. 1981 Nov;14(5):501–509. doi: 10.1128/jcm.14.5.501-509.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nickless G. G., Thorpe G. H., Kricka L. J., Whitehead T. P., Wells L. J., Ala F. A. A rapid chemiluminescent enzyme-linked immunosorbent assay for cytomegalovirus immunoglobulin G antibodies using instant photographic film. J Virol Methods. 1985 Dec;12(3-4):313–321. doi: 10.1016/0166-0934(85)90142-9. [DOI] [PubMed] [Google Scholar]
- Pronovost A. D., Baumgarten A., Hsiung G. D. Sensitive chemiluminescent enzyme-linked immunosorbent assay for quantification of human immunoglobulin G and detection of herpes simplex virus. J Clin Microbiol. 1981 Jan;13(1):97–101. doi: 10.1128/jcm.13.1.97-101.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thorpe G. H., Haggart R., Kricka L. J., Whitehead T. P. Enhanced luminescent enzyme immunoassays for rubella antibody, immunoglobulin E and digoxin. Biochem Biophys Res Commun. 1984 Mar 15;119(2):481–487. doi: 10.1016/s0006-291x(84)80274-0. [DOI] [PubMed] [Google Scholar]
- Thorpe G. H., Whitehead T. P., Penn R., Kricka L. J. Photographic monitoring of enhanced luminescent immunoassays. Clin Chem. 1984 May;30(5):806–807. [PubMed] [Google Scholar]