Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1979 Sep;10(3):317–321. doi: 10.1128/jcm.10.3.317-321.1979

Enzyme-linked fluorescence assay: Ultrasensitive solid-phase assay for detection of human rotavirus.

R H Yolken, P J Stopa
PMCID: PMC273160  PMID: 226564

Abstract

Enzyme-linked immunosorbent assay (ELISA) has proven to be a useful assay system for the direct detection of infectious agents. However, when the usual color-producing substrates are employed, relatively large amounts of substrate must be hydrolyzed by the bound enzyme before detection can be achieved. We attempted to improve the sensitivity of ELISA by utilizing a substrate that yields a fluorescent product on enzyme action. The enzyme-linked fluorescence assay (ELFA) based on this principle was approximately 100 times more sensitive than the corresponding ELISA or radioimmunoassay for the detection of human rotavirus in a standard stool suspension. In addition, the ELFA for human rotavirus was capable of detecting antigen in six specimens that were negative by ELISA. Five of these specimens were obtained late in the course of confirmed rotavirus infections. ELFA provides a simple, reliable, ultrasensitive method for the rapid detection of viral antigen.

Full text

PDF
317

Selected References

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

  1. Almeida J. D., Deinhardt F., Zuckerman A. J. Letter: Virus-like particles in hepatitis-A-positive faecal extracts. Lancet. 1974 Nov 2;2(7888):1083–1084. doi: 10.1016/s0140-6736(74)92188-6. [DOI] [PubMed] [Google Scholar]
  2. Broome C. V., Cherry W. B., Winn W. C., Jr, MacPherson B. R. Rapid diagnosis of Legionnaires' disease by direct immunofluorescent staining. Ann Intern Med. 1979 Jan;90(1):1–4. doi: 10.7326/0003-4819-90-1-1. [DOI] [PubMed] [Google Scholar]
  3. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol. 1972 Jul;109(1):129–135. [PubMed] [Google Scholar]
  4. Fernley H. N., Walker P. G. Kinetic behaviour of calf-intestinal alkaline phosphatase with 4-methylumbelliferyl phosphate. Biochem J. 1965 Oct;97(1):95–103. doi: 10.1042/bj0970095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gardner P. S., McQuillin J. Application of immunofluorescent antibody technique in rapid diagnosis of respiratory syncytial virus infection. Br Med J. 1968 Aug 10;3(5614):340–343. doi: 10.1136/bmj.3.5614.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Totterdell B. M., Chrystie I. L., Banatvala J. E. Rotavirus infections in a maternity unit. Arch Dis Child. 1976 Dec;51(12):924–928. doi: 10.1136/adc.51.12.924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Voller A., Bidwell D. E., Bartlett A. Enzyme immunoassays in diagnostic medicine. Theory and practice. Bull World Health Organ. 1976;53(1):55–65. [PMC free article] [PubMed] [Google Scholar]
  10. Walsh J. H., Yalow R., Berson S. A. Detection of Australia antigen and antibody by means of radioimmunoassay techniques. J Infect Dis. 1970 May;121(5):550–554. doi: 10.1093/infdis/121.5.550. [DOI] [PubMed] [Google Scholar]
  11. Waters M. D., Summer G. K., Hill H. D. Studies on the stability of 3-O-methyl fluorescein phosphate. Biochim Biophys Acta. 1968 Jun 4;159(2):420–422. doi: 10.1016/0005-2744(68)90096-x. [DOI] [PubMed] [Google Scholar]
  12. Wisdom G. B. Enzyme-immunoassay. Clin Chem. 1976 Aug;22(8):1243–1255. [PubMed] [Google Scholar]
  13. Yolken R. H., Barbour B., Wyatt R. G., Kalica A. R., Kapikian A. Z., Chanock R. M. Enzyme-linked immunosorbent assay for identification of rotaviruses from different animal species. Science. 1978 Jul 21;201(4352):259–262. doi: 10.1126/science.208150. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. 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]
  17. Yolken R., Wyatt R. G., Kapikian A. Z. ELISA for rotavirus. Lancet. 1977 Oct 15;2(8042):819–819. doi: 10.1016/s0140-6736(77)90750-4. [DOI] [PubMed] [Google Scholar]
  18. von Bonsdorff C. H., Hovi T., Mäkelä P., Mörttinen A. Rotavirus infections in adults in association with acute gastroenteritis. J Med Virol. 1978;2(1):21–28. doi: 10.1002/jmv.1890020105. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES