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. 1992 Nov 1;89(21):10011–10015. doi: 10.1073/pnas.89.21.10011

Diagnosis of hepatitis C virus (HCV) infection using an immunodominant chimeric polyprotein to capture circulating antibodies: reevaluation of the role of HCV in liver disease.

D Y Chien 1, Q L Choo 1, A Tabrizi 1, C Kuo 1, J McFarland 1, K Berger 1, C Lee 1, J R Shuster 1, T Nguyen 1, D L Moyer 1, et al.
PMCID: PMC50267  PMID: 1279666

Abstract

Structural and nonstructural regions of the HCV-encoded polyprotein have been expressed in recombinant yeast, bacteria, or insect cells and used to capture and measure reactive antibodies circulating in different individuals. The putative nucleocapsid protein (C) and nonstructural proteins 3-5 (NS3-NS5) were found to contain the most immunodominant epitopes. The NS3, NS4, and C regions were expressed in yeast in the form of a fused, chimeric polyprotein (C25) and a capture assay for reactive antibody was developed. This anti-C25 assay detects all previously identified HCV-seropositive cases and provides a substantially more sensitive diagnostic for both acute and chronic HCV infections than the current anti-C100-3 (NS4) assay. Anti-C25 was detected more frequently than anti-C100-3 in chronic, transfusion-associated non-A, non-B hepatitis patients from the United States (95% vs. 71%) and Japan (98% vs. 82%), in cryptogenic cirrhosis patients from the United States (62% vs. 28%), and in hepatitis B surface antigen-negative cases of hepatocellular carcinoma from Japan (83% vs. 63%). These data indicate that HCV has a greater role in these liver diseases than was previously thought. In volunteer United States blood donors sampled following the introduction of anti-C100-3 screening, the prevalence of anti-C25 and anti-C100-3 was 0.5% and 0.08%, respectively.

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Selected References

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  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. Alter M. J., Hadler S. C., Judson F. N., Mares A., Alexander W. J., Hu P. Y., Miller J. K., Moyer L. A., Fields H. A., Bradley D. W. Risk factors for acute non-A, non-B hepatitis in the United States and association with hepatitis C virus infection. JAMA. 1990 Nov 7;264(17):2231–2235. [PubMed] [Google Scholar]
  3. Austin H., Delzell E., Grufferman S., Levine R., Morrison A. S., Stolley P. D., Cole P. A case-control study of hepatocellular carcinoma and the hepatitis B virus, cigarette smoking, and alcohol consumption. Cancer Res. 1986 Feb;46(2):962–966. [PubMed] [Google Scholar]
  4. Chiba J., Ohba H., Matsuura Y., Watanabe Y., Katayama T., Kikuchi S., Saito I., Miyamura T. Serodiagnosis of hepatitis C virus (HCV) infection with an HCV core protein molecularly expressed by a recombinant baculovirus. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4641–4645. doi: 10.1073/pnas.88.11.4641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chlebowski R. T., Tong M., Weissman J., Block J. B., Ramming K. P., Weiner J. M., Bateman J. R., Chlebowski J. S. Hepatocellular carcinoma. Diagnostic and prognostic features in North American patients. Cancer. 1984 Jun 15;53(12):2701–2706. doi: 10.1002/1097-0142(19840615)53:12<2701::aid-cncr2820531224>3.0.co;2-b. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Choo Q. L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Coit D., Medina-Selby R., Barr P. J. Genetic organization and diversity of the hepatitis C virus. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2451–2455. doi: 10.1073/pnas.88.6.2451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Cousens L. S., Shuster J. R., Gallegos C., Ku L. L., Stempien M. M., Urdea M. S., Sanchez-Pescador R., Taylor A., Tekamp-Olson P. High level expression of proinsulin in the yeast, Saccharomyces cerevisiae. Gene. 1987;61(3):265–275. doi: 10.1016/0378-1119(87)90190-9. [DOI] [PubMed] [Google Scholar]
  10. Dawson G. J., Lesniewski R. R., Stewart J. L., Boardway K. M., Gutierrez R. A., Pendy L., Johnson R. G., Alcalde X., Rote K. V., Devare S. G. Detection of antibodies to hepatitis C virus in U.S. blood donors. J Clin Microbiol. 1991 Mar;29(3):551–556. doi: 10.1128/jcm.29.3.551-556.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Di Bisceglie A. M., Order S. E., Klein J. L., Waggoner J. G., Sjogren M. H., Kuo G., Houghton M., Choo Q. L., Hoofnagle J. H. The role of chronic viral hepatitis in hepatocellular carcinoma in the United States. Am J Gastroenterol. 1991 Mar;86(3):335–338. [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Hasan F., Jeffers L. J., De Medina M., Reddy K. R., Parker T., Schiff E. R., Houghton M., Choo Q. L., Kuo G. Hepatitis C-associated hepatocellular carcinoma. Hepatology. 1990 Sep;12(3 Pt 1):589–591. doi: 10.1002/hep.1840120323. [DOI] [PubMed] [Google Scholar]
  15. Hijikata M., Kato N., Ootsuyama Y., Nakagawa M., Shimotohno K. Gene mapping of the putative structural region of the hepatitis C virus genome by in vitro processing analysis. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5547–5551. doi: 10.1073/pnas.88.13.5547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hopf U., Möller B., Küther D., Stemerowicz R., Lobeck H., Lüdtke-Handjery A., Walter E., Blum H. E., Roggendorf M., Deinhardt F. Long-term follow-up of posttransfusion and sporadic chronic hepatitis non-A, non-B and frequency of circulating antibodies to hepatitis C virus (HCV). J Hepatol. 1990 Jan;10(1):69–76. doi: 10.1016/0168-8278(90)90075-3. [DOI] [PubMed] [Google Scholar]
  17. Houghton M., Weiner A., Han J., Kuo G., Choo Q. L. Molecular biology of the hepatitis C viruses: implications for diagnosis, development and control of viral disease. Hepatology. 1991 Aug;14(2):381–388. [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. 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]
  21. 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]
  22. Nasoff M. S., Zebedee S. L., Inchauspé G., Prince A. M. Identification of an immunodominant epitope within the capsid protein of hepatitis C virus. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5462–5466. doi: 10.1073/pnas.88.12.5462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Sansonno D., Dammacco F. Antibodies to hepatitis C virus in non-A, non-B post-transfusion and cryptogenetic chronic liver disease. Lancet. 1989 Sep 30;2(8666):798–799. doi: 10.1016/s0140-6736(89)90855-6. [DOI] [PubMed] [Google Scholar]
  26. Smith G. E., Ju G., Ericson B. L., Moschera J., Lahm H. W., Chizzonite R., Summers M. D. Modification and secretion of human interleukin 2 produced in insect cells by a baculovirus expression vector. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8404–8408. doi: 10.1073/pnas.82.24.8404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Takamizawa A., Mori C., Fuke I., Manabe S., Murakami S., Fujita J., Onishi E., Andoh T., Yoshida I., Okayama H. Structure and organization of the hepatitis C virus genome isolated from human carriers. J Virol. 1991 Mar;65(3):1105–1113. doi: 10.1128/jvi.65.3.1105-1113.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Van der Poel C. L., Cuypers H. T., Reesink H. W., Weiner A. J., Quan S., Di Nello R., Van Boven J. J., Winkel I., Mulder-Folkerts D., Exel-Oehlers P. J. Confirmation of hepatitis C virus infection by new four-antigen recombinant immunoblot assay. Lancet. 1991 Feb 9;337(8737):317–319. doi: 10.1016/0140-6736(91)90942-i. [DOI] [PubMed] [Google Scholar]
  31. Weiner A. J., Kuo G., Bradley D. W., Bonino F., Saracco G., Lee C., Rosenblatt J., Choo Q. L., Houghton M. Detection of hepatitis C viral sequences in non-A, non-B hepatitis. Lancet. 1990 Jan 6;335(8680):1–3. doi: 10.1016/0140-6736(90)90134-q. [DOI] [PubMed] [Google Scholar]
  32. Yarrish R. L., Werner B. G., Blumberg B. S. Association of hepatitis B virus infection with hepatocellular carcinoma in American patients. Int J Cancer. 1980 Dec 15;26(6):711–715. doi: 10.1002/ijc.2910260603. [DOI] [PubMed] [Google Scholar]
  33. van der Poel C. L., Reesink H. W., Lelie P. N., Leentvaar-Kuypers A., Choo Q. L., Kuo G., Houghton M. Anti-hepatitis C antibodies and non-A, non-B post-transfusion hepatitis in The Netherlands. Lancet. 1989 Aug 5;2(8658):297–298. doi: 10.1016/s0140-6736(89)90486-8. [DOI] [PubMed] [Google Scholar]
  34. van der Poel C. L., Reesink H. W., Schaasberg W., Leentvaar-Kuypers A., Bakker E., Exel-Oehlers P. J., Lelie P. N. Infectivity of blood seropositive for hepatitis C virus antibodies. Lancet. 1990 Mar 10;335(8689):558–560. doi: 10.1016/0140-6736(90)90347-8. [DOI] [PubMed] [Google Scholar]

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