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. 1980 Jan-Feb;53(1):85–92.

Enzyme-linked immunosorbent assay (ELISA): a practical tool for rapid diagnosis of viruses and other infectious agents.

R H Yolken
PMCID: PMC2595839  PMID: 6990637

Abstract

Tissue culture techniques are inadequate to diagnose some viral infections. Thus, solid-phase immunoassays have been developed for the direct detection of viral antigens in clinical specimens. While radioimmunoassays (RIA) have attained widespread use, solid-phase enzyme-linked immunosorbent assays (ELISA) offer a number of advantages over RIA systems. ELISAs can be established with approximately the same sensitivity as radioimmunoassays without utilizing unstable, gamma-emitting isotopes. However, before ELISA systems can obtain widespread usage, a number of aspects of the test must be optimized. These include the preparation and use of reagents, the nature of the solid phase, the choice of enzyme, and the enzyme-antibody conjugation method. With the solving of these problems, ELISA should attain widespread usage for rapid diagnosis of a large number of infectious agents.

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Selected References

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

  1. Almeida J. D., Rubenstein D., Stott E. J. New antigen-antibody system in Australia-antigen-positive hepatitis. Lancet. 1971 Dec 4;2(7736):1225–1227. doi: 10.1016/s0140-6736(71)90543-5. [DOI] [PubMed] [Google Scholar]
  2. Avrameas S., Ternynck T. Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penetration. Immunochemistry. 1971 Dec;8(12):1175–1179. doi: 10.1016/0019-2791(71)90395-8. [DOI] [PubMed] [Google Scholar]
  3. Bishop R. F., Davidson G. P., Holmes I. H., Ruck B. J. Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis. Lancet. 1973 Dec 8;2(7841):1281–1283. doi: 10.1016/s0140-6736(73)92867-5. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Connor J. D., Barrett-Connor E. Infectious diarrheas. Pediatr Clin North Am. 1967 Feb;14(1):197–221. doi: 10.1016/s0031-3955(16)31951-4. [DOI] [PubMed] [Google Scholar]
  6. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 1971 Sep;8(9):871–874. doi: 10.1016/0019-2791(71)90454-x. [DOI] [PubMed] [Google Scholar]
  7. Feinstone S. M., Kapikian A. Z., Purceli R. H. Hepatitis A: detection by immune electron microscopy of a viruslike antigen associated with acute illness. Science. 1973 Dec 7;182(4116):1026–1028. doi: 10.1126/science.182.4116.1026. [DOI] [PubMed] [Google Scholar]
  8. Hösli P., Avrameas S., Ullmann A., Vogt E., Rodrigot M. Quantitative ultramicro-scale immunoenzymic method for measuring Ig antigenic determinants in single cells. Clin Chem. 1978 Aug;24(8):1325–1330. [PubMed] [Google Scholar]
  9. Kapikian A. Z., Wyatt R. G., Dolin R., Thornhill T. S., Kalica A. R., Chanock R. M. Visualization by immune electron microscopy of a 27-nm particle associated with acute infectious nonbacterial gastroenteritis. J Virol. 1972 Nov;10(5):1075–1081. doi: 10.1128/jvi.10.5.1075-1081.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kitagawa T., Aikawa T. Enzyme coupled immunoassay of insulin using a novel coupling reagent. J Biochem. 1976 Jan;79(1):233–236. doi: 10.1093/oxfordjournals.jbchem.a131053. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Park H. A new plastic receptacle for solid phase immunoassays. J Immunol Methods. 1978;20:349–355. doi: 10.1016/0022-1759(78)90270-3. [DOI] [PubMed] [Google Scholar]
  13. Prince A. M. An antigen detected in the blood during the incubation period of serum hepatitis. Proc Natl Acad Sci U S A. 1968 Jul;60(3):814–821. doi: 10.1073/pnas.60.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Purcell R. H., Gerin J. L., Almeida J. B., Holland P. V. Radioimmunoassay for the detection of the core of the Dane particle and antibody to it. Intervirology. 1974;2(4):231–243. doi: 10.1159/000149428. [DOI] [PubMed] [Google Scholar]
  15. Purcell R. H., Wong D. C., Alter H. J., Holland P. V. Microtiter solid-phase radioimmunoassay for hepatitis B antigen. Appl Microbiol. 1973 Oct;26(4):478–484. doi: 10.1128/am.26.4.478-484.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ternynck T., Avrameas S. Conjugation of p-benzoquinone treated enzymes with antibodies and Fab fragments. Immunochemistry. 1977 Nov-Dec;14(11-12):767–774. doi: 10.1016/0019-2791(77)90352-4. [DOI] [PubMed] [Google Scholar]
  17. Watanabe H., Holmes I. H. Filter paper solid-phase radioimmunoassay for human rotavirus surface immunoglobulins. J Clin Microbiol. 1977 Oct;6(4):319–324. doi: 10.1128/jcm.6.4.319-324.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Watanabe N., Niitsu Y., Ohtsuka S., Koseki J., Kohgo Y., Urushizaki I., Kato K., Ishikawa E. Enzyme immunoassay for human ferritin. Clin Chem. 1979 Jan;25(1):80–82. [PubMed] [Google Scholar]
  19. Wisdom G. B. Enzyme-immunoassay. Clin Chem. 1976 Aug;22(8):1243–1255. [PubMed] [Google Scholar]
  20. Wolters G., Kuijpers L., Kacaki J., Schuurs A. Solid-phase enzyme-immunoassay for detection of hepatitis B surface antigen. J Clin Pathol. 1976 Oct;29(10):873–879. doi: 10.1136/jcp.29.10.873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yolken R. H. ELISA: enzyme-linked immunosorbent assay. Hosp Pract. 1978 Dec;13(12):121–127. doi: 10.1080/21548331.1978.11707455. [DOI] [PubMed] [Google Scholar]
  22. Yolken R. H., Greenberg H. B., Merson M. H., Sack R. B., Kapikian A. Z. Enzyme-linked immunosorbent assay for detection of Escherichia coli heat-labile enterotoxin. J Clin Microbiol. 1977 Nov;6(5):439–444. doi: 10.1128/jcm.6.5.439-444.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Yolken R. H., Stopa P. J. Analysis of nonspecific reactions in enzyme-linked immunosorbent assay testing for human rotavirus. J Clin Microbiol. 1979 Nov;10(5):703–707. doi: 10.1128/jcm.10.5.703-707.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Ziola B. R., Matikainen M. T., Salmi A. Polystyrene balls as the solid-phase of a double-antibody radioimmunoassay for human serum albumin. J Immunol Methods. 1977;17(3-4):309–317. doi: 10.1016/0022-1759(77)90113-2. [DOI] [PubMed] [Google Scholar]
  28. von der Waart M., Snelting A., Cichy J., Wolters G., Schuurs A. Enzyme-immunoassay in diagnosis of hepatitis with emphasis on the detection of "e" antigen (HBeAg). J Med Virol. 1978;3(1):43–49. doi: 10.1002/jmv.1890030111. [DOI] [PubMed] [Google Scholar]

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