1887

Abstract

Current anti-hepatitis C virus (HCV) antibody screening immunoassays are routinely based on an indirect format. Although their use for anti-HCV antibody detection has achieved a very high specificity and sensitivity, false-positive results are still a problem especially among populations with a low prevalence of HCV infection. One strategy to obviate this problem is to adapt the assay from an indirect format to a double-antigen sandwich one to further improve its specificity. In this study, a double-antigen sandwich time-resolved immunofluorometric assay (DAS-TRIFMA) has been developed to detect total anti-HCV antibodies based on biotin–streptavidin interaction. For comparison, 1025 samples were analysed by the DAS-TRIFMA and three indirect anti-HCV antibody detection methods. For samples with discordant results, PCR-ELISA and Inno-LIA were employed as supplementary assays to analyse the presence of HCV antibodies. With regard to the 1025 clinical samples, the overall concordance between the DAS-TRIFMA and the three indirect methods was 99.41, 98.93 and 98.93 % for Ortho ELISA 3.0, WAT ELISA and I-TRIFMA, respectively. The specificity/sensitivity of the DAS-TRIFMA, Ortho HCV ELISA 3.0, WAT HCV ELISA and I-TRIFMA were 100/99.09, 99.34/98.18, 99.23/97.27 and 99.01 %/98.18 %, respectively. The DAS-TRIFMA was able to detect HCV antibodies at a concentration about 1/10 of that detectable by indirect methods. From the obtained results and their comparison, it is concluded that the DAS-TRIFMA is a more specific and reliable method for screening anti-HCV antibodies, and weakly positive S/Co values by the DAS-TRIFMA were more predictive of HCV infection than those by indirect methods.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.47835-0
2008-08-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jmm/57/8/947.html?itemId=/content/journal/jmm/10.1099/jmm.0.47835-0&mimeType=html&fmt=ahah

References

  1. Abdel-Hamid M., El-Daly M., El-Kafrawy S., Mikhail N., Strickland G. T., Fix A. D. 2002; Comparison of second- and third-generation enzyme immunoassays for detecting antibodies to hepatitis C virus. J Clin Microbiol 40:1656–1659 [CrossRef]
    [Google Scholar]
  2. Alter M. J., Kuhnert W. L., Finelli L. 2003; Guidelines for laboratory testing and result reporting of antibody to hepatitis C virus. MMWR 52: (RR-3), 1–16
    [Google Scholar]
  3. Ansari M. H. K., Omrani M. D. 2006; Evaluation of diagnostic value of Elisa method (EIA) & PCR in diagnosis of hepatitis C virus in hemodialysis patients. Hepatitis Monthly 6:19–23
    [Google Scholar]
  4. Bürgisser P., Simon F., Wernli M., Wüst T., Beya M. F., Frei P. C. 1996; Multi-center evaluation of new double-antigen sandwich enzyme immunoassay for measurement of anti-human immunodeficiency virus type 1 and type 2 antibodies. J Clin Microbiol 34:634–637
    [Google Scholar]
  5. Carreño V. 2006; Occult hepatitis C virus infection: a new form of hepatitis C. World J Gastroenterol 12:6922–6925
    [Google Scholar]
  6. Carreño V., Pardo M., López-Alcorocho J. M., Rodríguez-Iñigo E., Bartolomé J., Castillo I. 2006; Detection of hepatitis C virus (HCV) RNA in the liver of healthy, anti-HCV antibody-positive, serum HCV RNA-negative patients with normal alanine aminotransferase levels. J Infect Dis 194:53–60 [CrossRef]
    [Google Scholar]
  7. CDC 2000; Hepatitis C virus infection among firefighters, emergency medical technicians, and paramedics – selected locations, United States, 1991–2000. MMWR 49:660–665
    [Google Scholar]
  8. Chevaliez S., Pawlotsky J. M. 2006; Hepatitis C virus serologic and virologic tests and clinical diagnosis of HCV related liver disease. Int J Med Sci 3:35–40
    [Google Scholar]
  9. Colin C., Lanoir D., Touzet S., Meyaud-Kraemer L., Bailly F., Trepo C. 2001; Sensitivity and specificity of third-generation hepatitis C virus antibody detection assays: an analysis of the literature. J Viral Hepat 8:87–95 [CrossRef]
    [Google Scholar]
  10. Dufour D. R., Talastas M., Fernandez M. D. A., Harris B., Strader D. B., Seeff L. B. 2003a; Low-positive anti-hepatitis C virus enzyme immunoassay results: an important predictor of low likelihood of hepatitis C infection. Clin Chem 49:479–486 [CrossRef]
    [Google Scholar]
  11. Dufour D. R., Talastas M., Fernandez M. D. A., Harris B. 2003b; Chemiluminescence assay improves specificity of hepatitis C antibody detection. Clin Chem 49:940–944 [CrossRef]
    [Google Scholar]
  12. Hyams K. C., Riddle J., Rubertone M., Trump D. H., Mazzuchi A. J. 2001; Prevalence and incidence of hepatitis C virus infection in the US military: a seroepidemiologic survey of 21,000 troops. Am J Epidemiol 153:764–770 [CrossRef]
    [Google Scholar]
  13. Lok A. S., Gunaratnam N. T. 1997; Diagnosis of hepatitis C. Hepatology 26 (Suppl. 1):48S–56S [CrossRef]
    [Google Scholar]
  14. Radkowski M., Horban A., Gallegos-Orozco J. F., Pawelczyk A., Jablonska J., Wilkinson J., Adair D., Laskus T. 2005; Evidence for viral persistence in patients who test positive for anti-hepatitis C virus antibodies and have normal alanine aminotransferase levels. J Infect Dis 191:1730–1733 [CrossRef]
    [Google Scholar]
  15. Ren H. R., Zhuang H. 2005; The progress in studying the correlation of S/Co values of anti-hepatitis C virus enzyme immunoassay with the positive results of supplemental assays. Chin J Lab Med 8:1096–1099 in Chinese
    [Google Scholar]
  16. Richter S. S. 2002; Laboratory assays for diagnosis and management of hepatitis C virus infection. J Clin Microbiol 40:4407–4412 [CrossRef]
    [Google Scholar]
  17. Schmidt B. L., Edjlalipour M., Luger A. 2000; Comparative evaluation of nine different enzyme-linked immunosorbent assays for determination of antibodies against Treponema pallidum in patients with primary syphilis. J Clin Microbiol 38:1279–1282
    [Google Scholar]
  18. Wu F. B., Xu Y. Y., Xu T., Wang Y. Z., Han S. Q. 1999; Time-resolved fluorescence immunoassay of thyroxine in serum: immobilized antigen approach. Anal Biochem 276:171–176 [CrossRef]
    [Google Scholar]
  19. Wu F. B., Han S. Q., He Y. F. 2002; Time-resolved immunofluorometry of serum hTSH with enhanced sensitivity. J Immunoassay Immunochem 23:191–210 [CrossRef]
    [Google Scholar]
  20. Zylberberg H., Pol S. 1996; Reciprocal interaction between human immunodeficiency virus and hepatitis C infection. Clin Infect Dis 23:1117–1125 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.47835-0
Loading
/content/journal/jmm/10.1099/jmm.0.47835-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error