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. 1991 Jun 1;88(11):4641–4645. doi: 10.1073/pnas.88.11.4641

Serodiagnosis of hepatitis C virus (HCV) infection with an HCV core protein molecularly expressed by a recombinant baculovirus.

J Chiba 1, H Ohba 1, Y Matsuura 1, Y Watanabe 1, T Katayama 1, S Kikuchi 1, I Saito 1, T Miyamura 1
PMCID: PMC51721  PMID: 1905012

Abstract

An enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of hepatitis C virus (HCV) infection, using HCV core protein (p22) synthesized by a recombinant baculovirus. Among 58 clinically well-defined chronic non-A, non-B hepatitis (NANBH) patients, 49 (84.5%) were positive for p22 antibody (anti-p22), whereas 42 (72.4%) were positive for C100-3 antibody (anti-C100-3), as measured by the present assay using the HCV nonstructural protein as antigen. Thirty-nine patients (67.2%) had both antibodies. No significant level of anti-p22 was detected in sera of chronic hepatitis B patients or normal blood donors. In typical post-transfusion NANBH patients, anti-p22 could be detected at, or even before, the first alanine aminotransferase peak. Anti-p22 was also detected in blood donors who were previously shown to be involved in transmitting HCV but in whose serum anti-C100-3 was not detectable. The ELISA detecting antibody to the HCV core protein expressed and properly processed in animal cells will be useful for mass screening of donor blood as well as for early diagnosis of hepatitis C.

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Selected References

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

  1. Alter H. J., Purcell R. H., Shih J. W., Melpolder J. C., Houghton M., Choo Q. L., Kuo G. Detection of antibody to hepatitis C virus in prospectively followed transfusion recipients with acute and chronic non-A, non-B hepatitis. N Engl J Med. 1989 Nov 30;321(22):1494–1500. doi: 10.1056/NEJM198911303212202. [DOI] [PubMed] [Google Scholar]
  2. Choo Q. L., Kuo G., Weiner A. J., Overby L. R., Bradley D. W., Houghton M. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science. 1989 Apr 21;244(4902):359–362. doi: 10.1126/science.2523562. [DOI] [PubMed] [Google Scholar]
  3. Choo Q. L., Weiner A. J., Overby L. R., Kuo G., Houghton M., Bradley D. W. Hepatitis C virus: the major causative agent of viral non-A, non-B hepatitis. Br Med Bull. 1990 Apr;46(2):423–441. doi: 10.1093/oxfordjournals.bmb.a072408. [DOI] [PubMed] [Google Scholar]
  4. Colombo M., Kuo G., Choo Q. L., Donato M. F., Del Ninno E., Tommasini M. A., Dioguardi N., Houghton M. Prevalence of antibodies to hepatitis C virus in Italian patients with hepatocellular carcinoma. Lancet. 1989 Oct 28;2(8670):1006–1008. doi: 10.1016/s0140-6736(89)91016-7. [DOI] [PubMed] [Google Scholar]
  5. Davis G. L., Balart L. A., Schiff E. R., Lindsay K., Bodenheimer H. C., Jr, Perrillo R. P., Carey W., Jacobson I. M., Payne J., Dienstag J. L. Treatment of chronic hepatitis C with recombinant interferon alfa. A multicenter randomized, controlled trial. Hepatitis Interventional Therapy Group. N Engl J Med. 1989 Nov 30;321(22):1501–1506. doi: 10.1056/NEJM198911303212203. [DOI] [PubMed] [Google Scholar]
  6. Di Bisceglie A. M., Martin P., Kassianides C., Lisker-Melman M., Murray L., Waggoner J., Goodman Z., Banks S. M., Hoofnagle J. H. Recombinant interferon alfa therapy for chronic hepatitis C. A randomized, double-blind, placebo-controlled trial. N Engl J Med. 1989 Nov 30;321(22):1506–1510. doi: 10.1056/NEJM198911303212204. [DOI] [PubMed] [Google Scholar]
  7. Dienstag J. L. non-A, Non-B hepatitis. II. Experimental transmission, putative virus agents and markers, and prevention. Gastroenterology. 1983 Sep;85(3):743–768. [PubMed] [Google Scholar]
  8. Enomoto N., Takada A., Nakao T., Date T. There are two major types of hepatitis C virus in Japan. Biochem Biophys Res Commun. 1990 Aug 16;170(3):1021–1025. doi: 10.1016/0006-291x(90)90494-8. [DOI] [PubMed] [Google Scholar]
  9. Esteban J. I., Esteban R., Viladomiu L., López-Talavera J. C., González A., Hernández J. M., Roget M., Vargas V., Genescà J., Buti M. Hepatitis C virus antibodies among risk groups in Spain. Lancet. 1989 Aug 5;2(8658):294–297. doi: 10.1016/s0140-6736(89)90485-6. [DOI] [PubMed] [Google Scholar]
  10. Esteban J. I., González A., Hernández J. M., Viladomiu L., Sánchez C., López-Talavera J. C., Lucea D., Martin-Vega C., Vidal X., Esteban R. Evaluation of antibodies to hepatitis C virus in a study of transfusion-associated hepatitis. N Engl J Med. 1990 Oct 18;323(16):1107–1112. doi: 10.1056/NEJM199010183231605. [DOI] [PubMed] [Google Scholar]
  11. Katayama T., Kikuchi S., Tanaka Y., Saito I., Miyamura T., Choo Q. L., Houghton M., Kuo G. Blood screening for non-A, non-B hepatitis by hepatitis C virus antibody assay. Transfusion. 1990 May;30(4):374–376. doi: 10.1046/j.1537-2995.1990.30490273448.x. [DOI] [PubMed] [Google Scholar]
  12. Kato N., Hijikata M., Ootsuyama Y., Nakagawa M., Ohkoshi S., Sugimura T., Shimotohno K. Molecular cloning of the human hepatitis C virus genome from Japanese patients with non-A, non-B hepatitis. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9524–9528. doi: 10.1073/pnas.87.24.9524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kuo G., Choo Q. L., Alter H. J., Gitnick G. L., Redeker A. G., Purcell R. H., Miyamura T., Dienstag J. L., Alter M. J., Stevens C. E. An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis. Science. 1989 Apr 21;244(4902):362–364. doi: 10.1126/science.2496467. [DOI] [PubMed] [Google Scholar]
  14. Matsuura Y., Miyamoto M., Sato T., Morita C., Yasui K. Characterization of Japanese encephalitis virus envelope protein expressed by recombinant baculoviruses. Virology. 1989 Dec;173(2):674–682. doi: 10.1016/0042-6822(89)90580-1. [DOI] [PubMed] [Google Scholar]
  15. Miller R. H., Purcell R. H. Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2057–2061. doi: 10.1073/pnas.87.6.2057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miyamura T., Saito I., Katayama T., Kikuchi S., Tateda A., Houghton M., Choo Q. L., Kuo G. Detection of antibody against antigen expressed by molecularly cloned hepatitis C virus cDNA: application to diagnosis and blood screening for posttransfusion hepatitis. Proc Natl Acad Sci U S A. 1990 Feb;87(3):983–987. doi: 10.1073/pnas.87.3.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Muraiso K., Hijikata M., Ohkoshi S., Cho M. J., Kikuchi M., Kato N., Shimotohno K. A structural protein of hepatitis C virus expressed in E. coli facilitates accurate detection of hepatitis C virus. Biochem Biophys Res Commun. 1990 Oct 30;172(2):511–516. doi: 10.1016/0006-291x(90)90702-o. [DOI] [PubMed] [Google Scholar]
  18. Nishioka K., Watanabe J., Furuta S., Tanaka E., Iino S., Suzuki H., Tsuji T., Yano M., Kuo G., Choo Q. L. A high prevalence of antibody to the hepatitis C virus in patients with hepatocellular carcinoma in Japan. Cancer. 1991 Jan 15;67(2):429–433. doi: 10.1002/1097-0142(19910115)67:2<429::aid-cncr2820670218>3.0.co;2-#. [DOI] [PubMed] [Google Scholar]
  19. Ohkoshi S., Kojima H., Tawaraya H., Miyajima T., Kamimura T., Asakura H., Satoh A., Hirose S., Hijikata M., Kato N. Prevalence of antibody against non-A, non-B hepatitis virus in Japanese patients with hepatocellular carcinoma. Jpn J Cancer Res. 1990 Jun-Jul;81(6-7):550–553. doi: 10.1111/j.1349-7006.1990.tb02605.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Okamoto H., Okada S., Sugiyama Y., Yotsumoto S., Tanaka T., Yoshizawa H., Tsuda F., Miyakawa Y., Mayumi M. The 5'-terminal sequence of the hepatitis C virus genome. Jpn J Exp Med. 1990 Jun;60(3):167–177. [PubMed] [Google Scholar]
  21. Saito I., Miyamura T., Ohbayashi A., Harada H., Katayama T., Kikuchi S., Watanabe Y., Koi S., Onji M., Ohta Y. Hepatitis C virus infection is associated with the development of hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6547–6549. doi: 10.1073/pnas.87.17.6547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith G. E., Summers M. D., Fraser M. J. Production of human beta interferon in insect cells infected with a baculovirus expression vector. Mol Cell Biol. 1983 Dec;3(12):2156–2165. doi: 10.1128/mcb.3.12.2156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Takeuchi K., Boonmar S., Kubo Y., Katayama T., Harada H., Ohbayashi A., Choo Q. L., Kuo G., Houghton M., Saito I. Hepatitis C viral cDNA clones isolated from a healthy carrier donor implicated in post-transfusion non-A, non-B hepatitis. Gene. 1990 Jul 16;91(2):287–291. doi: 10.1016/0378-1119(90)90102-w. [DOI] [PubMed] [Google Scholar]
  24. Takeuchi K., Kubo Y., Boonmar S., Watanabe Y., Katayama T., Choo Q. L., Kuo G., Houghton M., Saito I., Miyamura T. Nucleotide sequence of core and envelope genes of the hepatitis C virus genome derived directly from human healthy carriers. Nucleic Acids Res. 1990 Aug 11;18(15):4626–4626. doi: 10.1093/nar/18.15.4626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Takeuchi K., Kubo Y., Boonmar S., Watanabe Y., Katayama T., Choo Q. L., Kuo G., Houghton M., Saito I., Miyamura T. The putative nucleocapsid and envelope protein genes of hepatitis C virus determined by comparison of the nucleotide sequences of two isolates derived from an experimentally infected chimpanzee and healthy human carriers. J Gen Virol. 1990 Dec;71(Pt 12):3027–3033. doi: 10.1099/0022-1317-71-12-3027. [DOI] [PubMed] [Google Scholar]
  26. Thotakura N. R., Bahl O. P. Enzymatic deglycosylation of glycoproteins. Methods Enzymol. 1987;138:350–359. doi: 10.1016/0076-6879(87)38030-9. [DOI] [PubMed] [Google Scholar]
  27. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. WROBLEWSKI F., LADUE J. S. Serum glutamic pyruvic transaminase in cardiac with hepatic disease. Proc Soc Exp Biol Med. 1956 Apr;91(4):569–571. doi: 10.3181/00379727-91-22330. [DOI] [PubMed] [Google Scholar]

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