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. 1979 Nov;10(5):703–707. doi: 10.1128/jcm.10.5.703-707.1979

Analysis of nonspecific reactions in enzyme-linked immunosorbent assay testing for human rotavirus.

R H Yolken, P J Stopa
PMCID: PMC273251  PMID: 232506

Abstract

Solid-phase enzyme immunoassays can be utilized to detect antigens directly in clinical specimens. However, a small number of stools which we tested for human rotavirus by enzyme-linked immunosorbent assay (ELISA) were found to have nonspecific activity in the absence of rotaviral antigen. Similar nonspecific activity was found in eight of eight sera which contained rheumatoid factor. This nonspecific activity was markedly reduced by pretreatment of the specimens with reducing agents, normal goat serum, and anti-human immunoglobulin M (IgM). Thus, it is likely that these specimens contain an IgM antibody capable of reacting nonspecifically with the other components of the assay. Although pretreatment with the mild reducing agent N-acetylcysteine markedly reduced this nonspecific activity, such treatment did not reduce the specific ELISA activity due to rotavirus. Other treatments did produce a reduction in specific activity. Thus pretreatment with N-acetylcysteine offers a practical means to increase the specificity of ELISA systems without reducing their sensitivity.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Banatvala J. E., Best J. M., Kennedy E. A., Smith E. E., Spence M. E. A serological method for demonstrating recent infection by rubella virus. Br Med J. 1967 Jul 29;3(5560):285–286. doi: 10.1136/bmj.3.5560.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birch C. J., Lewis F. A., Kennett M. L., Homola M., Pritchard H., Gust I. D. A study of the prevalence of rotavirus infection in children with gastroenteritis admitted to an infectious diseases hospital. J Med Virol. 1977;1(1):69–77. doi: 10.1002/jmv.1890010109. [DOI] [PubMed] [Google Scholar]
  3. Carson D. A., Lawrance S., Catalano M. A., Vaughan J. H., Abraham G. Radioimmunoassay of IgG and IgM rheumatoid factors reacting with human IgG. J Immunol. 1977 Jul;119(1):295–300. [PubMed] [Google Scholar]
  4. Chao R. K., Fishaut M., Schwartzman J. D., McIntosh K. Detection of respiratory syncytial virus in nasal secretions from infants by enzyme-linked immunosorbent assay. J Infect Dis. 1979 Apr;139(4):483–486. doi: 10.1093/infdis/139.4.483. [DOI] [PubMed] [Google Scholar]
  5. Clem T. R., Yolken R. H. Practical colorimeter for direct measurement of microplates in enzyme immunoassay systems. J Clin Microbiol. 1978 Jan;7(1):55–58. doi: 10.1128/jcm.7.1.55-58.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cukor G., Berry M. K., Blacklow N. R. Simplified radioimmunoassay for detection of human rotavirus in stools. J Infect Dis. 1978 Dec;138(6):906–910. doi: 10.1093/infdis/138.6.906. [DOI] [PubMed] [Google Scholar]
  7. Ellens D. J., de Leeuw P. W. Detection of infantile gastroenteritis virus (rotavirus) by ELISA. Lancet. 1977 Jun 25;1(8026):1363–1364. doi: 10.1016/s0140-6736(77)92574-0. [DOI] [PubMed] [Google Scholar]
  8. Greenberg H. B., Wyatt R. G., Valdesuso J., Kalica A. R., London W. T., Chanock R. M., Kapikian A. Z. Solid-phase microtiter radioimmunoassay for detection of the Norwalk strain of acute nonbacterial, epidemic gastroenteritis virus and its antibodies. J Med Virol. 1978;2(2):97–108. doi: 10.1002/jmv.1890020204. [DOI] [PubMed] [Google Scholar]
  9. Johansson M. E., Bergquist N. R., Grandien M. Antibodies to calf serum as a cause of unwanted reaction in immunofluorescence tests. J Immunol Methods. 1976;11(3-4):265–272. doi: 10.1016/0022-1759(76)90119-8. [DOI] [PubMed] [Google Scholar]
  10. Kalica A. R., Purcell R. H., Sereno M. M., Wyatt R. G., Kim H. W., Chanock R. M., Kapikian A. Z. A microtiter solid phase radioimmunoassay for detection of the human reovirus-like agent in stools. J Immunol. 1977 Apr;118(4):1275–1279. [PubMed] [Google Scholar]
  11. Kapikian A. Z., Kim H. W., Wyatt R. G., Cline W. L., Arrobio J. O., Brandt C. D., Rodriguez W. J., Sack D. A., Chanock R. M., Parrott R. H. Human reovirus-like agent as the major pathogen associated with "winter" gastroenteritis in hospitalized infants and young children. N Engl J Med. 1976 Apr 29;294(18):965–972. doi: 10.1056/NEJM197604292941801. [DOI] [PubMed] [Google Scholar]
  12. Kapikian A. Z., Kim H. W., Wyatt R. G., Rodriguez W. J., Ross S., Cline W. L., Parrott R. H., Chanock R. M. Reoviruslike agent in stools: association with infantile diarrhea and development of serologic tests. Science. 1974 Sep 20;185(4156):1049–1053. doi: 10.1126/science.185.4156.1049. [DOI] [PubMed] [Google Scholar]
  13. Konno T., Suzuki H., Imai A., Ishida N. Reovirus-like agent in acute epidemic gastroenteritis in Japanese infants: fecal shedding and serologic response. J Infect Dis. 1977 Feb;135(2):259–266. doi: 10.1093/infdis/135.2.259. [DOI] [PubMed] [Google Scholar]
  14. Middleton P. J., Holdaway M. D., Petric M., Szymanski M. T., Tam J. S. Solid-phase radioimmunoassay for the detection of rotavirus. Infect Immun. 1977 May;16(2):439–444. doi: 10.1128/iai.16.2.439-444.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Middleton P. J., Petric M., Hewitt C. M., Szymanski M. T., Tam J. S. Counter-immunoelectro-osmophoresis for the detection of infantile gastroenteritis virus (orbi-group) antigen and antibody. J Clin Pathol. 1976 Mar;29(3):191–197. doi: 10.1136/jcp.29.3.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. PETERSON E. T., WALFORD R. L., FIGUEROA W. G., CHISHOLM R. Ox cell hemolysins in infectious mononucleosis and in other diseases. Am J Med. 1956 Aug;21(2):193–199. doi: 10.1016/0002-9343(56)90052-3. [DOI] [PubMed] [Google Scholar]
  17. Purcell R. H., Wong D. C., Moritsugu Y., Dienstag J. L., Routenberg J. A., Boggs J. D. A microtiter solid-phase radioimmunoassay for hepatitis A antigen and antibody. J Immunol. 1976 Feb;116(2):349–356. [PubMed] [Google Scholar]
  18. Shirodaria P. V., Fraser K. B., Stanford F. Secondary fluorescent staining of virus antigens by rheumatoid factor and fluorescein-conjugated anti-IgM. Ann Rheum Dis. 1973 Jan;32(1):53–57. doi: 10.1136/ard.32.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wisdom G. B. Enzyme-immunoassay. Clin Chem. 1976 Aug;22(8):1243–1255. [PubMed] [Google Scholar]
  20. Yolken R. H., Kim H. W., Clem T., Wyatt R. G., Kalica A. R., Chanock R. M., Kapikian A. Z. Enzyme-linked immunosorbent assay (ELISA) for detection of human reovirus-like agent of infantile gastroenteritis. Lancet. 1977 Aug 6;2(8032):263–267. doi: 10.1016/s0140-6736(77)90951-5. [DOI] [PubMed] [Google Scholar]
  21. Yolken R. H., Wyatt R. G., Barbour B. A., Kim H. W., Kapikian A. Z., Chanock R. M. Measurement of rotavirus antibody by an enzyme-linked immunosorbent assay blocking assay. J Clin Microbiol. 1978 Sep;8(3):283–287. doi: 10.1128/jcm.8.3.283-287.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yolken R. H., Wyatt R. G., Kalica A. R., Kim H. W., Brandt C. D., Parrott R. H., Kapikian A. Z., Chanock R. M. Use of a free viral immunofluorescence assay to detect human reovirus-like agent in human stools. Infect Immun. 1977 May;16(2):467–470. doi: 10.1128/iai.16.2.467-470.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yolken R. H., Wyatt R. G., Zissis G., Brandt C. D., Rodriguez W. J., Kim H. W., Parrott R. H., Urrutia J. J., Mata L., Greenberg H. B. Epidemiology of human rotavirus Types 1 and 2 as studied by enzyme-linked immunosorbent assay. N Engl J Med. 1978 Nov 23;299(21):1156–1161. doi: 10.1056/NEJM197811232992103. [DOI] [PubMed] [Google Scholar]

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