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. 1976 Mar;13(3):898–908. doi: 10.1128/iai.13.3.898-908.1976

Purification of hepatitis A antigen from feces and detection of antigen and antibody by immune adherence hemagglutination.

Y Moritsugu, J L Dienstag, J Valdesuso, D C Wong, J Wagner, J A Routenberg, R H Purcell
PMCID: PMC420693  PMID: 178597

Abstract

Hepatitis A antigen (HA Ag) was purified from feces collected during acute illness from patients with naturally occurring viral hepatitis, type A. Positive fecal specimens were identified by immune electron microscopy, but for detection of HA Agduring purification immune adherence hemagglutination (IAHA) and microtiter solid-phase radioimmunoassay were used. Isopycnic banding in cesium chloride, rate-zonal separation in sucrose, and preparative zonal electrophoresis were used in various combinations for successive purification, and the purified antigen was successfully used in a test for antibody by IAHA. Seronconversions to HA Ag were demonstrated by IAHA in 20 instances of hepatitis A virus infection, but in none of six cases of type B hepatitis or three cases of post-transfusion hepatitis unrelated to heaptitis A or B viruses, nor in two individuals without hepatitis. In addition, the temporal pattern of antibody development during type A hepatitis was studied in serial sera from an experimentally infected chimpanzee. Antibody titers by IAHA correlated well with antibody ratings determined by immune electron microscopy.

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Selected References

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

  1. Boggs J. D., Melnick J. L., Conrad M. E., Felsher B. F. Viral hepatitis. Clinical and tissue culture studies. JAMA. 1970 Nov 9;214(6):1041–1046. doi: 10.1001/jama.214.6.1041. [DOI] [PubMed] [Google Scholar]
  2. Deinhardt F., Holmes A. W., Capps R. B., Popper H. Studies on the transmission of human viral hepatitis to marmoset monkeys. I. Transmission of disease, serial passages, and description of liver lesions. J Exp Med. 1967 Apr 1;125(4):673–688. doi: 10.1084/jem.125.4.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dienstag J. L., Feinstone S. M., Purcell R. H., Hoofnagle J. H., Barker L. F., London W. T., Popper H., Peterson J. M., Kapikian A. Z. Experimental infection of chimpanzees with hepatitis A virus. J Infect Dis. 1975 Nov;132(5):532–545. doi: 10.1093/infdis/132.5.532. [DOI] [PubMed] [Google Scholar]
  4. Dienstag J. L., Routenberg J. A., Purcell R. H., Hooper R. R., Harrison W. O. Foodhandler-associated outbreak of hepatitis type A. An immune electron microscopic study. Ann Intern Med. 1975 Nov;83(5):647–650. doi: 10.7326/0003-4819-83-5-647. [DOI] [PubMed] [Google Scholar]
  5. Feinstone S. M., Kapikian A. Z., Gerin J. L., Purcell R. H. Buoyant density of the hepatitis A virus-like particle in cesium chloride. J Virol. 1974 Jun;13(6):1412–1414. doi: 10.1128/jvi.13.6.1412-1414.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Gravelle C. R., Hornbeck C. L., Maynard J. E., Schable C. A., Cook E. H., Bradley D. W. Hepatitis A: report of a common-source outbreak with recovery of a possible etiologic agent. II. Laboratory studies. J Infect Dis. 1975 Feb;131(2):167–171. doi: 10.1093/infdis/131.2.167. [DOI] [PubMed] [Google Scholar]
  8. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  9. Hollinger F. B., Bradley D. W., Maynard J. E., Dreesman G. R., Melnick J. L. Detection of hepatitis A viral antigen by radioimmunoassay. J Immunol. 1975 Nov;115(5):1464–1466. [PubMed] [Google Scholar]
  10. Holmes A. W., Deinhardt F., Wolfe L., Froesner G., Paterson D., Casto B., Conrad M. E. Specific neutralization of human hepatitis type A in marmoset monkeys. Nature. 1973 Jun 15;243(5407):419–420. doi: 10.1038/243419a0. [DOI] [PubMed] [Google Scholar]
  11. Holmes A. W., Wolfe L., Rosenblate H., Deinhardt F. Hepatitis in marmosets: induction of disease with coded specimens from a human volunteer study. Science. 1969 Aug 22;165(3895):816–817. doi: 10.1126/science.165.3895.816. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Knodell R. G., Conrad M. E., Dienstag J. L., Bell C. J. Etiological spectrum of post-transfusion hepatitis. Gastroenterology. 1975 Dec;69(6):1278–1285. [PubMed] [Google Scholar]
  14. Krugman S., Friedman H., Lattimer C. Viral hepatitis, type A. Identification by specific complement fixation and immune adherence tests. N Engl J Med. 1975 May 29;292(22):1141–1143. doi: 10.1056/NEJM197505292922201. [DOI] [PubMed] [Google Scholar]
  15. Krugman S., Giles J. P., Hammond J. Infectious hepatitis. Evidence for two distinctive clinical, epidemiological, and immunological types of infection. JAMA. 1967 May 1;200(5):365–373. doi: 10.1001/jama.200.5.365. [DOI] [PubMed] [Google Scholar]
  16. Lander J. J., Alter H. J., Purcell R. H. Frequency of antibody to hepatitis-associated antigen as measured by a new radioimmunoassay technique. J Immunol. 1971 May;106(5):1166–1171. [PubMed] [Google Scholar]
  17. Locarnini S. A., Ferris A. A., Stott A. C., Gust I. D. The relationship between a 27-nm virus-like particle and hepatitis A as demonstrated by immune electron microscopy. Intervirology. 1974;4(2):110–118. doi: 10.1159/000149849. [DOI] [PubMed] [Google Scholar]
  18. Mascoli C. C., Ittensohn O. L., Villarejos V. M., Arguedas J. A., Provost P. J., Hilleman M. R. Recovery of hepatitis agents in the marmoset from human cases occurring in Costa Rica. Proc Soc Exp Biol Med. 1973 Jan;142(1):276–282. doi: 10.3181/00379727-142-37005. [DOI] [PubMed] [Google Scholar]
  19. Matthew E. B., Dietzman D. E., Madden D. L., Newman S. J., Sever J. L., Nagler B., Bouton S. M., Rostafinski M. A major epidmeic of infectious hepatitis in an institution for the mentally retarded. Am J Epidemiol. 1973 Sep;98(3):199–215. doi: 10.1093/oxfordjournals.aje.a121549. [DOI] [PubMed] [Google Scholar]
  20. Mayumi M., Okochi K., Nishioka K. Detection of Australia antigen by means of immune adherence haemagglutination test. Vox Sang. 1971 Feb;20(2):178–181. doi: 10.1111/j.1423-0410.1971.tb00549.x. [DOI] [PubMed] [Google Scholar]
  21. Miller W. J., Provost P. J., McAleer W. J., Ittensohn O. L., Villarejos V. M., Hilleman M. R. Specific immune adherence assay for human hepatitis A antibody application to diagnostic and epidemiologic investigations. Proc Soc Exp Biol Med. 1975 May;149(1):254–261. doi: 10.3181/00379727-149-38783. [DOI] [PubMed] [Google Scholar]
  22. Morse L. J., Bryan J. A., Hurley J. P., Murphy J. F., O'Brien T. F., Wacker W. E. The Holy Cross college football team hepatitis outbreak. JAMA. 1972 Feb;219(6):706–708. [PubMed] [Google Scholar]
  23. Provost P. J., Ittensohn O. L., Villarejos V. M., Arguedas J. A., Hilleman M. R. Etiologic relationship of marmoset-propagated CR326 hepatitis A virus to hepatitis in man. Proc Soc Exp Biol Med. 1973 Apr;142(4):1257–1267. doi: 10.3181/00379727-142-37220. [DOI] [PubMed] [Google Scholar]
  24. Provost P. J., Ittensohn O. L., Villarejos V. M., Hilleman M. R. A specific complement-fixation test for human hepatitis a employing CR326 virus antigen. Diagnosis and epidemiology. Proc Soc Exp Biol Med. 1975 Apr;148(4):962–969. doi: 10.3181/00379727-148-38669. [DOI] [PubMed] [Google Scholar]
  25. Provost P. J., Wolanski B. S., Miller W. J., Ittensohn O. L., McAleer W. J., Hilleman M. R. Physical, chemical and morphologic dimensions of human hepatitis A virus strain CR326 (38578). Proc Soc Exp Biol Med. 1975 Feb;148(2):532–539. doi: 10.3181/00379727-148-38578. [DOI] [PubMed] [Google Scholar]
  26. Purcell R. H., Dienstag J. L., Feinstone S. M., Kapikian A. Z. Relationship of hepatitis A antigen to viral hepatitis. Am J Med Sci. 1975 Jul-Aug;270(1):61–71. doi: 10.1097/00000441-197507000-00010. [DOI] [PubMed] [Google Scholar]
  27. 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]

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