Abstract
A neutralization enzyme immunoassay (N-EIA) was developed for the detection of antibody titer rises in sera of patients infected with influenza A (H3N2) virus. In this N-EIA, a selected strain of influenza A (H3N2) virus was added to monolayers of LLC-MK2 cells in microtiter plates. After 24 h, the replicated virus could be demonstrated with a virus-specific enzyme-labeled monoclonal antibody. Preincubation of the influenza virus with convalescent-phase sera of patients infected with influenza A (H3N2) virus resulted 1 day later in decreased absorbance values that could be used for calculation of neutralization titers. From use of paired serum samples from 10 patients with a history of flu-like symptoms, the results obtained with N-EIA correlated well (r = 0.83) with those of the standard hemagglutination inhibition test.
Full text
PDF![987](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5930/267167/6051ae290de2/jcm00004-0143.png)
![988](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5930/267167/d8fa4cf514c1/jcm00004-0144.png)
![989](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5930/267167/edcc08787931/jcm00004-0145.png)
![990](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5930/267167/f894becb24a7/jcm00004-0146.png)
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson L. J., Hierholzer J. C., Bingham P. G., Stone Y. O. Microneutralization test for respiratory syncytial virus based on an enzyme immunoassay. J Clin Microbiol. 1985 Dec;22(6):1050–1052. doi: 10.1128/jcm.22.6.1050-1052.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Frank A. L., Puck J., Hughes B. J., Cate T. R. Microneutralization test for influenza A and B and parainfluenza 1 and 2 viruses that uses continuous cell lines and fresh serum enhancement. J Clin Microbiol. 1980 Sep;12(3):426–432. doi: 10.1128/jcm.12.3.426-432.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gitelman A. K., Kaverin N. V., Kharitonenkov I. G., Rudneva I. A., Sklyanskaya E. L., Zhdanov V. M. Dissociation of the haemagglutination inhibition and the infectivity neutralization in the reactions of influenza A/USSR/90/77 (H1N1) virus variants with monoclonal antibodies. J Gen Virol. 1986 Oct;67(Pt 10):2247–2251. doi: 10.1099/0022-1317-67-10-2247. [DOI] [PubMed] [Google Scholar]
- Gross P. A., Davis A. E. Neutralization test in influenza: use in individuals without hemagglutination inhibition antibody. J Clin Microbiol. 1979 Sep;10(3):382–384. doi: 10.1128/jcm.10.3.382-384.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harder T. C., Klusmeyer K., Frey H. R., Orvell C., Liess B. Intertypic differentiation and detection of intratypic variants among canine and phocid morbillivirus isolates by kinetic neutralization using a novel immunoplaque assay. J Virol Methods. 1993 Jan;41(1):77–92. doi: 10.1016/0166-0934(93)90164-m. [DOI] [PubMed] [Google Scholar]
- Harmsen T., Jongerius M. C., van der Zwan C. W., Plantinga A. D., Kraaijeveld C. A., Berbers G. A. Comparison of a neutralization enzyme immunoassay and an enzyme-linked immunosorbent assay for evaluation of immune status of children vaccinated for mumps. J Clin Microbiol. 1992 Aug;30(8):2139–2144. doi: 10.1128/jcm.30.8.2139-2144.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirst G. K. THE AGGLUTINATION OF RED CELLS BY ALLANTOIC FLUID OF CHICK EMBRYOS INFECTED WITH INFLUENZA VIRUS. Science. 1941 Jul 4;94(2427):22–23. doi: 10.1126/science.94.2427.22. [DOI] [PubMed] [Google Scholar]
- Lazarowitz S. G., Choppin P. W. Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the hemagglutinin polypeptide. Virology. 1975 Dec;68(2):440–454. doi: 10.1016/0042-6822(75)90285-8. [DOI] [PubMed] [Google Scholar]
- Lazarowitz S. G., Compans R. W., Choppin P. W. Influenza virus structural and nonstructural proteins in infected cells and their plasma membranes. Virology. 1971 Dec;46(3):830–843. doi: 10.1016/0042-6822(71)90084-5. [DOI] [PubMed] [Google Scholar]
- Nakane P. K., Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem. 1974 Dec;22(12):1084–1091. doi: 10.1177/22.12.1084. [DOI] [PubMed] [Google Scholar]
- Okuno Y., Tanaka K., Baba K., Maeda A., Kunita N., Ueda S. Rapid focus reduction neutralization test of influenza A and B viruses in microtiter system. J Clin Microbiol. 1990 Jun;28(6):1308–1313. doi: 10.1128/jcm.28.6.1308-1313.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Outlaw M. C., Dimmock N. J. Insights into neutralization of animal viruses gained from study of influenza virus. Epidemiol Infect. 1991 Apr;106(2):205–220. doi: 10.1017/s0950268800048354. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tannock G. A., Paul J. A., Herd R., Barry R. D., Reid A. L., Hensley M. J., Gillett R. S., Gillett S. M., Lawrance P., Henry R. L. Improved colorimetric assay for detecting influenza B virus neutralizing antibody responses to vaccination and infection. J Clin Microbiol. 1989 Mar;27(3):524–528. doi: 10.1128/jcm.27.3.524-528.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vlaspolder F., Harmsen T., van Veenendaal D., Kraaijeveld C. A., Snippe H. Application of immunoassay of encephalomyocarditis virus in cell culture with enzyme-labeled virus-specific monoclonal antibodies for rapid detection of virus, neutralizing antibodies, and interferon. J Clin Microbiol. 1988 Dec;26(12):2593–2597. doi: 10.1128/jcm.26.12.2593-2597.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Watanabe H., Mackenzie J. S. The detection of influenza A virus antigens in cultured cells by enzyme-linked immunosorbent assay. Arch Virol. 1981;67(1):31–43. doi: 10.1007/BF01314599. [DOI] [PubMed] [Google Scholar]
- de Jong J. C., de Ronde-Verloop F. M., Veenendaal-van Herk T. M., Weijers T. F., Bijlsma K., Osterhaus A. D. Antigenic heterogeneity within influenza A (H3N2) virus strains. Bull World Health Organ. 1988;66(1):47–55. [PMC free article] [PubMed] [Google Scholar]
- van Tiel F. H., Boere W. A., Vinjé J., Harmsen T., Benaissa-Trouw B. J., Kraaijeveld C. A., Snippe H. Detection of Semliki Forest virus in cell culture by use of an enzyme immunoassay with peroxidase-labeled monoclonal antibodies specific for glycoproteins E1 and E2. J Clin Microbiol. 1984 Sep;20(3):387–390. doi: 10.1128/jcm.20.3.387-390.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Tiel F. H., Harmsen T., Wagenaar M., Boere W. A., Kraaijeveld C. A., Snippe H. Rapid determination of neutralizing antibodies to Semliki Forest virus in serum by enzyme immunoassay in cell culture with virus-specific monoclonal antibodies. J Clin Microbiol. 1986 Oct;24(4):665–668. doi: 10.1128/jcm.24.4.665-668.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]