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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Jul;35(7):1734–1739. doi: 10.1128/jcm.35.7.1734-1739.1997

Serological determination of hepatitis C virus genotype: comparison with a standardized genotyping assay.

J M Pawlotsky 1, L Prescott 1, P Simmonds 1, C Pellet 1, P Laurent-Puig 1, C Labonne 1, F Darthuy 1, J Remire 1, J Duval 1, C Buffet 1, J P Etienne 1, D Dhumeaux 1, E Dussaix 1
PMCID: PMC229831  PMID: 9196183

Abstract

In patients with chronic hepatitis C, determination of hepatitis C virus (HCV) genotype could be routinely run in the future to tailor treatment schedules. The suitabilities of two versions of a serological, so-called serotyping assay (Murex HCV Serotyping Assay version 1-3 [SA1-3] and Murex HCV Serotyping Assay version 1-6 [SA1-6]; Murex Diagnostics Ltd.), based on the detection of genotype-specific antibodies directed to epitopes encoded by the NS4 region of the genome, for the routine determination of HCV genotypes were studied. The results were compared with those of a molecular biology-based genotyping method (HCV Line Probe Assay [INNO-LiPA HCV]; Innogenetics S.A.), based on hybridization of PCR products onto genotype-specific probes designed in the 5' noncoding region of the genome, obtained with pretreatment serum samples from 88 patients with chronic hepatitis C eligible for interferon therapy. Definitive genotyping was performed by sequence analysis of three regions of the viral genome in all samples with discrepant typing results found among at least two of the three assays studied. In all instances, sequence analysis confirmed the result of the INNO-LiPA HCV test. The sensitivity of SA1-3 was 75% relative to the results obtained by the genotyping assay. The results were concordant with those of genotyping for 92% of the samples typeable by SA1-3. The sensitivity of SA1-6 was 89% relative to the results obtained by the genotyping assay. The results were concordant with those of genotyping for 94% of the samples typeable by SA1-6. Overall, SA1-6 had increased sensitivity relative to SA1-3 but remained less sensitive than the genotyping assay on the basis of PCR amplification of HCV RNA. Cross-reactivities between different HCV genotypes could be responsible for the mistyping of 8 (SA1-3) and 6% (SA1-6) of the samples. Subtyping of 1a and 1b is still not possible with the existing peptides, but discriminating between subtypes may not be necessary for routine use.

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Selected References

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  1. Bhattacherjee V., Prescott L. E., Pike I., Rodgers B., Bell H., El-Zayadi A. R., Kew M. C., Conradie J., Lin C. K., Marsden H. Use of NS-4 peptides to identify type-specific antibody to hepatitis C virus genotypes 1, 2, 3, 4, 5 and 6. J Gen Virol. 1995 Jul;76(Pt 7):1737–1748. doi: 10.1099/0022-1317-76-7-1737. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Davidson F., Simmonds P., Ferguson J. C., Jarvis L. M., Dow B. C., Follett E. A., Seed C. R., Krusius T., Lin C., Medgyesi G. A. Survey of major genotypes and subtypes of hepatitis C virus using RFLP of sequences amplified from the 5' non-coding region. J Gen Virol. 1995 May;76(Pt 5):1197–1204. doi: 10.1099/0022-1317-76-5-1197. [DOI] [PubMed] [Google Scholar]
  4. Dixit V., Quan S., Martin P., Larson D., Brezina M., DiNello R., Sra K., Lau J. Y., Chien D., Kolberg J. Evaluation of a novel serotyping system for hepatitis C virus: strong correlation with standard genotyping methodologies. J Clin Microbiol. 1995 Nov;33(11):2978–2983. doi: 10.1128/jcm.33.11.2978-2983.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Féray C., Gigou M., Samuel D., Paradis V., Mishiro S., Maertens G., Reynés M., Okamoto H., Bismuth H., Bréchot C. Influence of the genotypes of hepatitis C virus on the severity of recurrent liver disease after liver transplantation. Gastroenterology. 1995 Apr;108(4):1088–1096. doi: 10.1016/0016-5085(95)90207-4. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Mahaney K., Tedeschi V., Maertens G., Di Bisceglie A. M., Vergalla J., Hoofnagle J. H., Sallie R. Genotypic analysis of hepatitis C virus in American patients. Hepatology. 1994 Dec;20(6):1405–1411. doi: 10.1002/hep.1840200605. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Mellor J., Holmes E. C., Jarvis L. M., Yap P. L., Simmonds P. Investigation of the pattern of hepatitis C virus sequence diversity in different geographical regions: implications for virus classification. The International HCV Collaborative Study Group. J Gen Virol. 1995 Oct;76(Pt 10):2493–2507. doi: 10.1099/0022-1317-76-10-2493. [DOI] [PubMed] [Google Scholar]
  10. Mondelli M. U., Cerino A., Bono F., Cividini A., Maccabruni A., Aricò M., Malfitano A., Barbarini G., Piazza V., Minoli L. Hepatitis C virus (HCV) core serotypes in chronic HCV infection. J Clin Microbiol. 1994 Oct;32(10):2523–2527. doi: 10.1128/jcm.32.10.2523-2527.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nousbaum J. B., Pol S., Nalpas B., Landais P., Berthelot P., Bréchot C. Hepatitis C virus type 1b (II) infection in France and Italy. Collaborative Study Group. Ann Intern Med. 1995 Feb 1;122(3):161–168. doi: 10.7326/0003-4819-122-3-199502010-00001. [DOI] [PubMed] [Google Scholar]
  12. Okamoto H., Mishiro S., Tokita H., Tsuda F., Miyakawa Y., Mayumi M. Superinfection of chimpanzees carrying hepatitis C virus of genotype II/1b with that of genotype III/2a or I/1a. Hepatology. 1994 Nov;20(5):1131–1136. [PubMed] [Google Scholar]
  13. 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]
  14. Pawlotsky J. M., Roudot-Thoraval F., Bastie A., Darthuy F., Rémiré J., Métreau J. M., Zafrani E. S., Duval J., Dhumeaux D. Factors affecting treatment responses to interferon-alpha in chronic hepatitis C. J Infect Dis. 1996 Jul;174(1):1–7. doi: 10.1093/infdis/174.1.1. [DOI] [PubMed] [Google Scholar]
  15. Pawlotsky J. M., Roudot-Thoraval F., Bastie A., Pellet C., Duval J., Dhumeaux D. Are hepatitis C virus genotypes 1a and 1b so different? Hepatology. 1996 May;23(5):1289–1291. doi: 10.1002/hep.510230556. [DOI] [PubMed] [Google Scholar]
  16. Pawlotsky J. M., Roudot-Thoraval F., Pellet C., Aumont P., Darthuy F., Remire J., Duval J., Dhumeaux D. Influence of hepatitis C virus (HCV) genotypes on HCV recombinant immunoblot assay patterns. J Clin Microbiol. 1995 May;33(5):1357–1359. doi: 10.1128/jcm.33.5.1357-1359.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Simmonds P., Rose K. A., Graham S., Chan S. W., McOmish F., Dow B. C., Follett E. A., Yap P. L., Marsden H. Mapping of serotype-specific, immunodominant epitopes in the NS-4 region of hepatitis C virus (HCV): use of type-specific peptides to serologically differentiate infections with HCV types 1, 2, and 3. J Clin Microbiol. 1993 Jun;31(6):1493–1503. doi: 10.1128/jcm.31.6.1493-1503.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Stuyver L., Wyseur A., van Arnhem W., Lunel F., Laurent-Puig P., Pawlotsky J. M., Kleter B., Bassit L., Nkengasong J., van Doorn L. J. Hepatitis C virus genotyping by means of 5'-UR/core line probe assays and molecular analysis of untypeable samples. Virus Res. 1995 Oct;38(2-3):137–157. doi: 10.1016/0168-1702(95)00052-r. [DOI] [PubMed] [Google Scholar]
  22. Stuyver L., van Arnhem W., Wyseur A., Hernandez F., Delaporte E., Maertens G. Classification of hepatitis C viruses based on phylogenetic analysis of the envelope 1 and nonstructural 5B regions and identification of five additional subtypes. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):10134–10138. doi: 10.1073/pnas.91.21.10134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tanaka T., Tsukiyama-Kohara K., Yamaguchi K., Yagi S., Tanaka S., Hasegawa A., Ohta Y., Hattori N., Kohara M. Significance of specific antibody assay for genotyping of hepatitis C virus. Hepatology. 1994 Jun;19(6):1347–1353. [PubMed] [Google Scholar]
  24. Tsukiyama-Kohara K., Yamaguchi K., Maki N., Ohta Y., Miki K., Mizokami M., Ohba K., Tanaka S., Hattori N., Nomoto A. Antigenicities of Group I and II hepatitis C virus polypeptides--molecular basis of diagnosis. Virology. 1993 Feb;192(2):430–437. doi: 10.1006/viro.1993.1058. [DOI] [PubMed] [Google Scholar]

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