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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Oct;34(10):2382–2385. doi: 10.1128/jcm.34.10.2382-2385.1996

Discordant results from hepatitis C virus genotyping by procedures based on amplification of different genomic regions.

P Toniutto 1, M Pirisi 1, S G Tisminetzky 1, C Fabris 1, E Chinellato 1, M Gerotto 1, E Falleti 1, P Ferroni 1, T Lombardelli 1, E Bartoli 1, F Baralle 1
PMCID: PMC229274  PMID: 8880484

Abstract

We compared the results of genotyping hepatitis C virus (HCV) either by PCR amplification of the core region or by hybridization of PCR-amplified products of the 5' untranslated region (5'UTR assay). Serum samples from 144 Italian anti-HCV-positive patients (106 drug abusers and 38 patients with chronic viral liver disease but no history of drug abuse) were studied. The original core region assay described by Okamoto et al. (H. Y. Okamoto, Y. Sugiyama, S. Okada, K. Kurai, Y. Akahane, Y. Sugai, T. Tanaka, K. Sato, F. Tsuda, Y. Miyakawa, and M. Mayumi, J. Gen. Virol. 73:673-679, 1992) allowed genotyping of 75 of 144 samples. A modified version of Widell et al. (A. Widell, S. Shev, S. Månsson, Y.-Y. Zhang, U. Foberg, G. Norkrans, A. Frydén, O. Weiland, J. Kurkus, and E. Nordenfelt, J. Med. Virol. 44:272-279, 1994) allowed genotyping of 11 of 79 samples (50 of 79 samples remained unclassified by the method of Okamoto et al. In contrast, all 144 samples were genotyped by the 5'UTR assay. Forty-six of 75 (61 percent) of the samples genotyped by the method of Okamoto et al. and 10 of 11 (91 percent) of the samples genotyped by the method of Widell et al. had results consistent with those obtained by the 5'UTR assay. According to the results of direct sequencing, the method of Okamoto et al. erroneously classified seven samples as having mixed infections. In conclusion, HCV genotyping seems more reliable when it is performed by the 5'UTR assay than by either of two core region assays. The major advantage provided by the 5'UTR assay is a much lower proportion of negative or indeterminate results in younger patients with histories of drug abuse or infection by genotypes other than HCV type 1.

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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. Bréchot C., Kremsdorf D. Genetic variation of the hepatitis C virus (HCV) genome: random events or a clinically relevant issue? J Hepatol. 1993 Mar;17(3):265–268. doi: 10.1016/s0168-8278(05)80203-x. [DOI] [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. Dusheiko G., Schmilovitz-Weiss H., Brown D., McOmish F., Yap P. L., Sherlock S., McIntyre N., Simmonds P. Hepatitis C virus genotypes: an investigation of type-specific differences in geographic origin and disease. Hepatology. 1994 Jan;19(1):13–18. [PubMed] [Google Scholar]
  5. Hayashi N., Higashi H., Kaminaka K., Sugimoto H., Esumi M., Komatsu K., Hayashi K., Sugitani M., Suzuki K., Tadao O. Molecular cloning and heterogeneity of the human hepatitis C virus (HCV) genome. J Hepatol. 1993;17 (Suppl 3):S94–107. doi: 10.1016/s0168-8278(05)80432-5. [DOI] [PubMed] [Google Scholar]
  6. Honda M., Kaneko S., Sakai A., Unoura M., Murakami S., Kobayashi K. Degree of diversity of hepatitis C virus quasispecies and progression of liver disease. Hepatology. 1994 Nov;20(5):1144–1151. [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. 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]
  10. 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]
  11. Pawlotsky J. M., Tsakiris L., Roudot-Thoraval F., Pellet C., Stuyver L., Duval J., Dhumeaux D. Relationship between hepatitis C virus genotypes and sources of infection in patients with chronic hepatitis C. J Infect Dis. 1995 Jun;171(6):1607–1610. doi: 10.1093/infdis/171.6.1607. [DOI] [PubMed] [Google Scholar]
  12. Silini E., Bono F., Cividini A., Cerino A., Bruno S., Rossi S., Belloni G., Brugnetti B., Civardi E., Salvaneschi L. Differential distribution of hepatitis C virus genotypes in patients with and without liver function abnormalities. Hepatology. 1995 Feb;21(2):285–290. [PubMed] [Google Scholar]
  13. Silini E., Bono F., Cividini A., Cerino A., Maccabruni A., Tinelli C., Bruno S., Bellobuono A., Mondelli M. Molecular epidemiology of hepatitis C virus infection among intravenous drug users. J Hepatol. 1995 Jun;22(6):691–695. doi: 10.1016/0168-8278(95)80225-8. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Simmonds P. Variability of hepatitis C virus. Hepatology. 1995 Feb;21(2):570–583. doi: 10.1002/hep.1840210243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Tsubota A., Chayama K., Ikeda K., Yasuji A., Koida I., Saitoh S., Hashimoto M., Iwasaki S., Kobayashi M., Hiromitsu K. Factors predictive of response to interferon-alpha therapy in hepatitis C virus infection. Hepatology. 1994 May;19(5):1088–1094. [PubMed] [Google Scholar]
  17. Widell A., Shev S., Månsson S., Zhang Y. Y., Foberg U., Norkrans G., Frydén A., Weiland O., Kurkus J., Nordenfelt E. Genotyping of hepatitis C virus isolates by a modified polymerase chain reaction assay using type specific primers: epidemiological applications. J Med Virol. 1994 Nov;44(3):272–279. doi: 10.1002/jmv.1890440311. [DOI] [PubMed] [Google Scholar]
  18. Yamada M., Kakumu S., Yoshioka K., Higashi Y., Tanaka K., Ishikawa T., Takayanagi M. Hepatitis C virus genotypes are not responsible for development of serious liver disease. Dig Dis Sci. 1994 Feb;39(2):234–239. doi: 10.1007/BF02090191. [DOI] [PubMed] [Google Scholar]

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