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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Jan;35(1):201–207. doi: 10.1128/jcm.35.1.201-207.1997

New hepatitis C virus (HCV) genotyping system that allows for identification of HCV genotypes 1a, 1b, 2a, 2b, 3a, 3b, 4, 5a, and 6a.

O Ohno 1, M Mizokami 1, R R Wu 1, M G Saleh 1, K Ohba 1, E Orito 1, M Mukaide 1, R Williams 1, J Y Lau 1
PMCID: PMC229539  PMID: 8968908

Abstract

Recent studies have focused on whether different hepatitis C virus (HCV) genotypes are associated with different profiles of pathogenicity, infectivity, and response to antiviral therapy. The establishment of a simple and precise genotyping system for HCV is essential to address these issues. A new genotyping system based on PCR of the core region with genotype-specific PCR primers for the determination of HCV genotypes 1a, 1b, 2a, 2b, 3a, 3b, 4, 5a, and 6a was developed. A total of 607 samples (379 from Japan, 63 from the United States, 53 from Korea, 35 from Taiwan, 32 from China, 20 from Hong Kong, 15 from Australia, 6 from Egypt, 3 from Bangladesh, and 1 from South Africa) were tested by both the assay of Okamoto et al. (H. Okamoto, Y. Sugiyama, S. Okada, K. Kurai, Y. Akahane, Y. Sugai, T. Tanaka, K. Sato, F. Tsuda, Y. Miyamura, and M. Mayumi, J. Gen. Virol. 73:673-679, 1992) and this new genotyping system. Comparison of the results showed concordant results for 539 samples (88.8%). Of the 68 samples with discordant results, the nucleotide sequences of the HCV isolates were determined in 23, and their genotypes were determined by molecular evolutionary analysis. In all 23 samples, the assignment of genotype by our new genotyping system was correct. This genotyping system may be useful for large-scale determination of HCV genotypes in clinical studies.

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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. Bukh J., Purcell R. H., Miller R. H. At least 12 genotypes of hepatitis C virus predicted by sequence analysis of the putative E1 gene of isolates collected worldwide. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8234–8238. doi: 10.1073/pnas.90.17.8234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bukh J., Purcell R. H., Miller R. H. Sequence analysis of the core gene of 14 hepatitis C virus genotypes. Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8239–8243. doi: 10.1073/pnas.91.17.8239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chayama K., Tsubota A., Arase Y., Saitoh S., Koida I., Ikeda K., Matsumoto T., Kobayashi M., Iwasaki S., Koyama S. Genotypic subtyping of hepatitis C virus. J Gastroenterol Hepatol. 1993 Mar-Apr;8(2):150–156. doi: 10.1111/j.1440-1746.1993.tb01507.x. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. 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]
  8. Giannini C., Thiers V., Nousbaum J. B., Stuyver L., Maertens G., Bréchot C. Comparative analysis of two assays for genotyping hepatitis C virus based on genotype-specific primers or probes. J Hepatol. 1995 Sep;23(3):246–253. [PubMed] [Google Scholar]
  9. Gojobori T., Ishii K., Nei M. Estimation of average number of nucleotide substitutions when the rate of substitution varies with nucleotide. J Mol Evol. 1982;18(6):414–423. doi: 10.1007/BF01840889. [DOI] [PubMed] [Google Scholar]
  10. Inchauspe G., Zebedee S., Lee D. H., Sugitani M., Nasoff M., Prince A. M. Genomic structure of the human prototype strain H of hepatitis C virus: comparison with American and Japanese isolates. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10292–10296. doi: 10.1073/pnas.88.22.10292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kanai K., Kako M., Okamoto H. HCV genotypes in chronic hepatitis C and response to interferon. Lancet. 1992 Jun 20;339(8808):1543–1543. doi: 10.1016/0140-6736(92)91311-u. [DOI] [PubMed] [Google Scholar]
  12. Kaneko S., Unoura M., Kobayashi K., Kuno K., Murakami S., Hattori N. Detection of serum hepatitis C virus RNA. Lancet. 1990 Apr 21;335(8695):976–976. doi: 10.1016/0140-6736(90)91042-9. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Kleter G. E., van Doorn L. J., Stuyver L., Maertens G., Brouwer J. T., Schalm S. W., Heijtink R. A., Quint W. G. Rapid genotyping of hepatitis C virus RNA-isolates obtained from patients residing in western Europe. J Med Virol. 1995 Sep;47(1):35–42. doi: 10.1002/jmv.1890470108. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Lau J. Y., Mizokami M., Kolberg J. A., Davis G. L., Prescott L. E., Ohno T., Perrillo R. P., Lindsay K. L., Gish R. G., Qian K. P. Application of six hepatitis C virus genotyping systems to sera from chronic hepatitis C patients in the United States. J Infect Dis. 1995 Feb;171(2):281–289. doi: 10.1093/infdis/171.2.281. [DOI] [PubMed] [Google Scholar]
  17. McOmish F., Chan S. W., Dow B. C., Gillon J., Frame W. D., Crawford R. J., Yap P. L., Follett E. A., Simmonds P. Detection of three types of hepatitis C virus in blood donors: investigation of type-specific differences in serologic reactivity and rate of alanine aminotransferase abnormalities. Transfusion. 1993 Jan;33(1):7–13. doi: 10.1046/j.1537-2995.1993.33193142314.x. [DOI] [PubMed] [Google Scholar]
  18. Mita E., Hayashi N., Kanazawa Y., Hagiwara H., Ueda K., Kasahara A., Fusamoto H., Kamada T. Hepatitis C virus genotype and RNA titer in the progression of type C chronic liver disease. J Hepatol. 1994 Sep;21(3):468–473. doi: 10.1016/s0168-8278(05)80330-7. [DOI] [PubMed] [Google Scholar]
  19. Mizokami M., Gojobori T., Lau J. Y. Molecular evolutionary virology: its application to hepatitis C virus. Gastroenterology. 1994 Oct;107(4):1181–1182. doi: 10.1016/0016-5085(94)90245-3. [DOI] [PubMed] [Google Scholar]
  20. Mori S., Kato N., Yagyu A., Tanaka T., Ikeda Y., Petchclai B., Chiewsilp P., Kurimura T., Shimotohno K. A new type of hepatitis C virus in patients in Thailand. Biochem Biophys Res Commun. 1992 Feb 28;183(1):334–342. doi: 10.1016/0006-291x(92)91648-a. [DOI] [PubMed] [Google Scholar]
  21. Mosley J. W., Aach R. D., Hollinger F. B., Stevens C. E., Barbosa L. H., Nemo G. J., Holland P. V., Bancroft W. H., Zimmerman H. J., Kuo G. Non-A, non-B hepatitis and antibody to hepatitis C virus. JAMA. 1990 Jan 5;263(1):77–78. [PubMed] [Google Scholar]
  22. Nakao T., Enomoto N., Takada N., Takada A., Date T. Typing of hepatitis C virus genomes by restriction fragment length polymorphism. J Gen Virol. 1991 Sep;72(Pt 9):2105–2112. doi: 10.1099/0022-1317-72-9-2105. [DOI] [PubMed] [Google Scholar]
  23. Nei M., Gojobori T. Simple methods for estimating the numbers of synonymous and nonsynonymous nucleotide substitutions. Mol Biol Evol. 1986 Sep;3(5):418–426. doi: 10.1093/oxfordjournals.molbev.a040410. [DOI] [PubMed] [Google Scholar]
  24. Ogata N., Alter H. J., Miller R. H., Purcell R. H. Nucleotide sequence and mutation rate of the H strain of hepatitis C virus. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3392–3396. doi: 10.1073/pnas.88.8.3392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ohba K., Mizokami M., Ohno T., Suzuki K., Orito E., Ina Y., Lau J. Y., Gojobori T. Classification of hepatitis C virus into major types and subtypes based on molecular evolutionary analysis. Virus Res. 1995 May;36(2-3):201–214. doi: 10.1016/0168-1702(95)00007-d. [DOI] [PubMed] [Google Scholar]
  26. Ohno T., Mizokami M., Tibbs C. J., Ohba K., Suzuki K., Wu R. R., Nouri-Aria K. T., Williams R. New genotype of hepatitis C virus in South Africa. J Med Virol. 1994 Apr;42(4):409–413. doi: 10.1002/jmv.1890420414. [DOI] [PubMed] [Google Scholar]
  27. Ohno T., Mizokami M., Yamauchi M., Ohba K., Orito E., Wu R. R., Mizuno M., Sugihara K., Wakita T., Kakumu S. Genotype distribution in Nagoya and new genotype (genotype 3a) in Japanese patients with hepatitis C virus. J Gastroenterol. 1995 Apr;30(2):209–214. doi: 10.1007/BF02348667. [DOI] [PubMed] [Google Scholar]
  28. Okamoto H., Kojima M., Okada S., Yoshizawa H., Iizuka H., Tanaka T., Muchmore E. E., Peterson D. A., Ito Y., Mishiro S. Genetic drift of hepatitis C virus during an 8.2-year infection in a chimpanzee: variability and stability. Virology. 1992 Oct;190(2):894–899. doi: 10.1016/0042-6822(92)90933-g. [DOI] [PubMed] [Google Scholar]
  29. Okamoto H., Kurai K., Okada S., Yamamoto K., Lizuka H., Tanaka T., Fukuda S., Tsuda F., Mishiro S. Full-length sequence of a hepatitis C virus genome having poor homology to reported isolates: comparative study of four distinct genotypes. Virology. 1992 May;188(1):331–341. doi: 10.1016/0042-6822(92)90762-e. [DOI] [PubMed] [Google Scholar]
  30. Okamoto H., Okada S., Sugiyama Y., Kurai K., Iizuka H., Machida A., Miyakawa Y., Mayumi M. Nucleotide sequence of the genomic RNA of hepatitis C virus isolated from a human carrier: comparison with reported isolates for conserved and divergent regions. J Gen Virol. 1991 Nov;72(Pt 11):2697–2704. doi: 10.1099/0022-1317-72-11-2697. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Okamoto H., Sugiyama Y., Okada S., Kurai K., Akahane Y., Sugai Y., Tanaka T., Sato K., Tsuda F., Miyakawa Y. Typing hepatitis C virus by polymerase chain reaction with type-specific primers: application to clinical surveys and tracing infectious sources. J Gen Virol. 1992 Mar;73(Pt 3):673–679. doi: 10.1099/0022-1317-73-3-673. [DOI] [PubMed] [Google Scholar]
  33. Okamoto H., Tokita H., Sakamoto M., Horikita M., Kojima M., Iizuka H., Mishiro S. Characterization of the genomic sequence of type V (or 3a) hepatitis C virus isolates and PCR primers for specific detection. J Gen Virol. 1993 Nov;74(Pt 11):2385–2390. doi: 10.1099/0022-1317-74-11-2385. [DOI] [PubMed] [Google Scholar]
  34. Pozzato G., Moretti M., Franzin F., Crocè L. S., Tiribelli C., Masayu T., Kaneko S., Unoura M., Kobayashi K. Severity of liver disease with different hepatitis C viral clones. Lancet. 1991 Aug 24;338(8765):509–509. doi: 10.1016/0140-6736(91)90578-d. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  37. Simmonds P., Alberti A., Alter H. J., Bonino F., Bradley D. W., Brechot C., Brouwer J. T., Chan S. W., Chayama K., Chen D. S. A proposed system for the nomenclature of hepatitis C viral genotypes. Hepatology. 1994 May;19(5):1321–1324. [PubMed] [Google Scholar]
  38. Simmonds P., Holmes E. C., Cha T. A., Chan S. W., McOmish F., Irvine B., Beall E., Yap P. L., Kolberg J., Urdea M. S. Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region. J Gen Virol. 1993 Nov;74(Pt 11):2391–2399. doi: 10.1099/0022-1317-74-11-2391. [DOI] [PubMed] [Google Scholar]
  39. Stuyver L., Rossau R., Wyseur A., Duhamel M., Vanderborght B., Van Heuverswyn H., Maertens G. Typing of hepatitis C virus isolates and characterization of new subtypes using a line probe assay. J Gen Virol. 1993 Jun;74(Pt 6):1093–1102. doi: 10.1099/0022-1317-74-6-1093. [DOI] [PubMed] [Google Scholar]
  40. Yoshioka K., Kakumu S., Wakita T., Ishikawa T., Itoh Y., Takayanagi M., Higashi Y., Shibata M., Morishima T. Detection of hepatitis C virus by polymerase chain reaction and response to interferon-alpha therapy: relationship to genotypes of hepatitis C virus. Hepatology. 1992 Aug;16(2):293–299. doi: 10.1002/hep.1840160203. [DOI] [PubMed] [Google Scholar]

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