1887

Abstract

An ELISA was used to detect a protein derived from the core gene of the hepatitis C virus (HCV) in human plasma. The solid phase antibody in the assay was a murine monoclonal antibody against a synthetic peptide deduced from the putative core gene of HCV (residues 39 to 74). An enzyme-labelled affinity-purified human antibody directed at another region within the HCV core (residues 5 to 23) was the second antibody tracer. The ELISA had a sensitivity capable of detecting a few ng/ml of the HCV core polypeptide expressed in . Core antigen activity in plasma of infected hosts was detected after treatment of HCV RNA-rich fractions from buoyant density centrifugation with the detergent Tween 80. There was a direct correlation between core antigen ELISA values of a plasma fraction and intensities of polymerase chain reaction signals for HCV RNA. These observations are consistent with the proposal that the N-terminal sequence of the predicted polyprotein of HCV is a nucleocapsid protein, and that improved core antigen assays may correlate with viraemia.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-73-3-667
1992-03-01
2024-12-09
Loading full text...

Full text loading...

/deliver/fulltext/jgv/73/3/JV0730030667.html?itemId=/content/journal/jgv/10.1099/0022-1317-73-3-667&mimeType=html&fmt=ahah

References

  1. Almeida J. D., Rubenstein D., Stott E. J. 1971; New antigenantibody system in Australia-antigen-positive hepatitis. Lancet ii:1225–1227
    [Google Scholar]
  2. Bradley D. W., Maynard J. E., Popper H., Cook E. H., Ebert J. W., McCaustland K. A., Schable C. A., Fields H. A. 1983; Posttransfusion non-A, non-B hepatitis: physicochemical properties of two distinct agents. Journal of Infectious Diseases 148:254–265
    [Google Scholar]
  3. Choo Q. L., Kuo G., Weiner A. J., Overby L. R., Bradley D. W., Houghton M. 1989; Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359–362
    [Google Scholar]
  4. Choo Q. L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Coit D., Medina-Selby A., Barr P. J., Weiner A. J., Bradley D. W., Kuo G., Houghton M. 1991; Genetic organization and diversity of the hepatitis C virus. Proceedings of the National Academy of Sciences, U.S.A 88:2451–2455
    [Google Scholar]
  5. Dane D. S., Cameron C. H., Briggs M. 1970; Virus-like particles in serum of patients with Australia-antigen-associated hepatitis. Lancet i:695–698
    [Google Scholar]
  6. Hoofnagle J. H., Gerety R. J., Barker L. F. 1973; Antibody to hepatitis B virus core in man. Lancet ii:869–873
    [Google Scholar]
  7. Kato N., Huikata M., Ootsuyama Y., Nakagawa M., Ookoshi S., Sugimura T., Shimotohno K. 1990; Molecular cloning of the human hepatitis C virus genome from Japanese patients with non-A, non-B hepatitis. Proceedings of the National Academy of Sciences, U.S.A 87:9524–9528
    [Google Scholar]
  8. Kuo G., Choo Q. L., Alter H. J., Gitnik G. L., Redeker A. G., Purcell R. H., Miyamura T., Dienstag J. L., Alter M. J., Stevens C. E., Tegtmeier G. E., Bonino F., Colombo M., Lee W. S., Kuo C., Berger K., Shuster J. R., Overby L. R., Bradley D. W., Houghton M. 1989; An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis. Science 244:362–364
    [Google Scholar]
  9. Miller R. H., Purcell R. H. 1990; Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups. Proceedings of the National Academy of Sciences, U.S.A 87:2057–2061
    [Google Scholar]
  10. Miyamoto H., Okamoto H., Sato K., Tanaka T., Mishiro S. 1992; Extraordinarily low density of hepatitis C virus estimated by sucrose density gradient centrifugation and the polymerase chain reaction. Journal of General Virology 73:715–718
    [Google Scholar]
  11. Nakane P. K., Kawaoi A. 1974; Peroxidase labelled antibody: a new method of conjugation. Journal of Histochemistry and Cyto-chemistry 22:1084–1091
    [Google Scholar]
  12. Oi V. R., Herzenberg L. A. 1980; Immunoglobulin producing hybrid cell lines. In Selected Methods in Cellular Immunology p 351 Edited by Mishell B., Shiigi S. M. San Francisco: Freeman;
    [Google Scholar]
  13. Okamoto H., Okada S., Sugiyama Y., Yotsumoto S., Tanaka T., Yoshizawa H., Tsuda F., Miyakawa Y., Mayumi M. 1990a; The 5´terminal sequence of the hepatitis C virus genome. Japanese Journal of Experimental Medicine 60:167–177
    [Google Scholar]
  14. Okamoto H., Okada S., Sugiyama Y., Tanaka T., Sugai Y., Akahane Y., Machida A., Mishiro S., Yoshizawa H., Miyakawa Y., Mayumi M. 1990b; Detection of hepatitis C virus RNA by a two-stage polymerase chain reaction with two pairs of primers deduced from the 5´noncoding region. Japanese Journal of Experimental Medicine 60:215–222
    [Google Scholar]
  15. Okamoto H., Munekata E., Tsuda F., Takahashi K., Yotsumoto S., Tanaka T., Tachibana K., Akahane Y., Sugai Y., Miyakawa Y., Mayumi M. 1990c; Enzyme-linked immunosorbent assay for antibodies against the capsid protein of hepatitis C virus with a synthetic oligopeptide. Japanese Journal of Experimental Medicine 60:223–233
    [Google Scholar]
  16. Shikata T., Karasawa T., Abe K., Uzawa T., Suzuki H., Oda T., Imai M., Mayumi M., Moritsugu Y. 1977; Hepatitis Be antigen and infectivity of hepatitis B virus. Journal of Infectious Diseases 136:571–576
    [Google Scholar]
  17. Takamizawa A., Mori C., Fuke I., Manabe S., Murakami S., Fujita J., Onishi E., Andoh T., Yoshida I., Okayama H. 1991; Structure and organization of the hepatitis C virus genome isolated from human carriers. Journal of Virology 65:1105–1113
    [Google Scholar]
  18. Weiner A. J., Kuo G., Bradley D. W., Bonino F., Saracco G., Lee C., Rosenblatt J., Choo Q. L., Houghton M. 1990; Detection of hepatitis C viral sequences in non-A, non-B hepatitis. Lancet 3351–3
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-73-3-667
Loading
/content/journal/jgv/10.1099/0022-1317-73-3-667
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error