Evaluation of the performance of the EIAgen HCV test for detection of hepatitis C virus infection

https://doi.org/10.1016/j.jviromet.2009.08.008Get rights and content

Abstract

The EIAgen HCV test (Adaltis Inc., Montreal, Canada) is an enzyme immunoassay (EIA) for the detection of anti-hepatitis C virus (HCV) antibodies. This study compared the performance of this test side-by-side with the current Ortho HCV 3.0 Anti-HCV assay (Ortho-Clinical Diagnostics Inc., Johnson & Johnson Company, Raritan, NY, USA). Among 2559 specimens examined, 178 were true positives, 2376 were true negatives and 5 were indeterminate. The sensitivity of the EIAgen HCV test was 100%, versus 98.3% for the Ortho HCV test, while their respective specificities were 98.1% and 98.2%. The EIAgen HCV test gave a positive predictive value of 79.8% and a negative predictive value of 100%. Overall, the concordance of this test with the Ortho HCV test was 98.2%. Specimens from potentially interfering substances, such as sera from pregnant women, sera from patients with acute non-C hepatitis, autoimmune diseases, lipidemia, or from patients undergoing hemolysis, showed no interference with either EIA. An EIAgen HCV test signal-to-cut-off ratio of >5.9 would be highly predictive of a true-positive finding in these specimens. The EIAgen HCV test is well suited for screening blood and blood products in antibodies to HCV.

Introduction

Hepatitis C virus (HCV) infection is a major global healthcare problem. The World Health Organization (WHO) estimates that up to 3% of the world's population has been infected with HCV, equating to more than 170 million carriers of HCV worldwide. The infection is often asymptomatic, but the infection becomes persistent in more than 80% of infected people and can be associated with chronic hepatitis, cirrhosis or hepatocellular carcinomas (Di Bisceglie, 2000, Hoofnagle, 2002, Wei et al., 2002, Afdhal, 2004). Transmission is associated mainly with infected blood products or intravenous drug abuse, although other less common routes such as perinatal or sexual transmission have been described (Vandelli et al., 2004). HCV was first identified in 1989 using a recombinant DNA approach (Choo et al., 1989). Currently, routine diagnosis of HCV is based on detecting anti-HCV IgG antibodies in serum or plasma by enzyme immunoassay (EIA) (Centers for Disease Control and Prevention, 1991). Screening of blood and blood products for anti-HCV antibodies is an important element part of prevention. Infection is defined as repeated reactivity by a screening assay and confirmed reactivity of ≥1+ for at least two HCV bands by a recombinant immunoblot assay (RIBA), with or without the presence of detectable viral RNA (Allain, 2005). To increase the safety of blood products, many countries have now implemented donor testing of pooled or single blood samples for viral RNA using a nucleic acid amplification test (NAT) (Centers for Disease Control and Prevention, 1998). In addition to NAT screening, serological testing for HCV antibodies has improved in both sensitivity and specificity. Implementation of blood donation screening for anti-HCV antibodies by EIA has led to a marked decline in the risk of transfusion-transmitted hepatitis (Donahue et al., 1992).

The EIAgen HCV test (Adaltis Inc., Montreal, Canada) is a diagnostic tool using EIA for qualitative detection of anti-HCV IgG and IgM antibodies in serum and plasma. The Ortho HCV 3.0 Anti-HCV (Ortho HCV) assay (Ortho-Clinical Diagnostics Inc., Johnson & Johnson Company, Raritan, NY, USA) is also a microplate EIA used to detect qualitatively anti-HCV antibodies. This test has been widely used, and its clinical performance is well documented (Tobler et al., 2003). This study was undertaken to compare the performance of the two tests with regards to sensitivity and specificity. The supplemental tests applied were the Chiron RIBA HCV 3.0 strip immunoblot assay (RIBA 3.0; Chiron Corporation, Emeryville, CA, USA) and a commercial NAT (Beijing Hepatitis Reagent Research & Production Center, Beijing, China).

Section snippets

Materials

EIAgen HCV tests were obtained from Adaltis Inc. (Montreal, Canada). These use a two-step EIA for the detection of HCV infections in serum and plasma. Microplate wells are coated with HCV-specific antigens which capture anti-HCV antibodies in the sample. Upon completion of the assay, the development of color indicates the presence of HCV. The Ortho HCV tests were purchased from Ortho-Clinical Diagnostics Inc. (Johnson & Johnson Company, Raritan, NY, USA). Confirmatory assays were the RIBA 3.0

Overall results

The results of 2559 sera tested by the EIAgen HCV test and the Ortho HCV Test are summarized in Table 1. Fourteen specimens (0.6%) had discordant results and were tested further by supplemental assays, as shown in Table 2. Five samples were RIBA 3.0 indeterminate, 3 were positive and 6 were negative. Overall, testing by the EIAgen HCV test yielded 178 true positives, no false negatives, 2331 true negatives and 45 false positives. Testing with the Ortho HCV test yielded 175 true positives, 3

Discussion

The EIAgen HCV test is a diagnostic EIA intended for screening patients with HCV infection using qualitative detection of anti-HCV IgG and IgM. The Ortho HCV test is a commercial assay, also using EIA, for qualitative detection of anti-HCV antibodies. The performance of the EIAgen HCV test with the current Ortho HCV test was evaluated using a series of specimens collected in clinical settings.

The main criteria for an HCV screening assay are to attain the highest sensitivity possible in

Acknowledgments

This work was partially supported by grants from the Chinese Basic Research Foundation 973 (2005CB522902 and 2007CB512900), 863 program (2006AA02A410), NSFC (30571639), the National Science and Technology Key Project during the 11th Five-Year Plan Period (2008ZX10002-012 and 2008ZX10002-013) and Key Clinical Research Program of Ministry of Health.

References (23)

  • J.P. Allain

    Hepatitis C virus in blood donation

    Lancet

    (2005)
  • N.H. Afdhal

    The natural history of hepatitis C

    Semin. Liver Dis.

    (2004)
  • M.J. Alter et al.

    Guidelines for laboratory testing and result reporting of antibody to hepatitis C virus

    M. M. W. R.

    (2003)
  • S. Bar-Shany et al.

    False positive tests for anti-hepatitis C antibodies and the problem of notifying blood donors

    Int. J. Epidemiol.

    (1996)
  • M.P. Busch et al.

    Very low level viremia in HCV infectious unit missed by NAT

    Transfusion

    (2003)
  • Centers for Disease Control and Prevention

    Public health service inter-agency guidelines for screening donors of blood, plasma, organ, tissues and semen for evidence of hepatitis B and hepatitis C

    M. M. W. R.

    (1991)
  • Centers for Disease Control and Prevention, 1998. Recommendations for prevention and control of hepatitis C virus (HCV)...
  • Q.L. Choo et al.

    Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome

    Science

    (1989)
  • A.M. Di Bisceglie

    Natural history of hepatitis C: its impact on clinical management

    Hepatology

    (2000)
  • J.G. Donahue et al.

    The declining risk of post-transfusion hepatitis C virus infection

    N. Engl. J. Med.

    (1992)
  • S.C. Du et al.

    Utilization of uracil-DNA glycosylase for combining reverse transcription and anti-contamination with polymerase chain reaction in hepatitis C virus

    J. Peking Univ. [Health Sci.]

    (2007)
  • Cited by (6)

    • Association between HCV infection and diabetes type 2 in Egypt: Is it time to split up?

      2015, Annals of Epidemiology
      Citation Excerpt :

      A third generation Enzyme Immunoassay (ELISA), Adlatis EIAgen HCV Ab test (Adaltis Inc., Montreal, Canada) was used to test for antibodies against HCV. This test showed 100% of sensitivity and 98.1% of specificity, with a cut-off value (signal-to-cut-off ratio, S/CO) of 1.0 or more to be considered reactive for anti-HCV antibodies, whereas those S/CO less than 1.0 were considered nonreactive [27]. All positives with this test were confirmed by a Chemiluminescent Microplate Immunoassay (CIA) [24].

    • Double-antigen sandwich ELISA for the detection of anti-hepatitis C virus antibodies

      2011, Journal of Virological Methods
      Citation Excerpt :

      The results from 16 specimens with chronic hepatitis C suggested that the double-antigen sandwich ELISA can effectively detect HCV-specific IgM antibodies. Although a new kit, named EIAgen HCV, that was recently developed in Canada can detect both IgG and IgM (Rao et al., 2009), this assay still cannot prevent non-specific interference because HRP-labeled anti-human IgG and IgM are used as the secondary antibodies. Therefore, using the double-antigen sandwich ELISA to detect IgM antibodies is better than using an indirect assay in terms of specificity.

    • The threshold value of anti-HCV test in the diagnosis of HCV infection

      2012, Turkiye Klinikleri Journal of Medical Sciences
    1

    These authors contributed equally.

    View full text