Development of low cost peptide-based anti-hepatitis C virus screening and confirmatory assays: Comparison with commercially available tests

Abstract

Screening and confirmatory low cost reagent tests have been developed for detection of anti-hepatitis C virus (HCV). Assays are based on the use of specific synthetic peptides from several structural and non-structural viral proteins. The efficacy of the screening anti-HCV EIA-Spep assay was compared with both Abbott EIA 2.0 (Abbott Laboratories, North Chicago, IL) and Ortho EIA 2.0 (Ortho Diagnostic Systems, Raritan, NJ) anti-HCV detection kits and the confirmatory EIA-Cpep assay was compared with the Abbott Matrix anti-HCV confirmation test. In the EIA-Spep, a pool of 3 peptides was added to each well of a microtiter plate. In EIA-Cpep, each well was separately coated with 1 of 4 peptides and 1 recombinant protein. A total of 867 blood donor samples from Costa Rica tested simultaneously with the 3 screening assays yielded the same specificity and negative predictive values of > or =99.9% and 100%, respectively. A comparative study on voluntary blood donor samples from Honduras, Nicaragua, and El Salvador using the 2 anti-HCV confirmatory assays revealed different patterns that are 46% positive, 24% indeterminate, and 30% negative with the EIA-Cpep assay vs. 31% positive, 48% indeterminate, and 21% negative with the Matrix assay. A study of 71 patient samples from Costa Rica showed a higher correlation between initially reactive samples when analyzed by the Abbott and Ortho kits, than when the assay results were compared between the Abbott and EIA-Spep kits; the latter detected 7 and 15 non-reactive samples, respectively. These results could reflect the use of a similar antigen source for the 2 commercial assays. The presence of HCV RNA in a group of 29 samples analyzed was related to the simultaneous reactivity in all 3 screening assays. None of the discordant samples had detectable levels of HCV RNA. Economic difficulties for health care services in the developing countries of Central America have prevented implementation of routine anti-HCV blood donor screening tests. This is likely to be the primary reason for uncontrolled dissemination of HCV, and the lack of identification of potential high risk groups. Alternative low cost reagents developed locally as described in this article could be a useful tool in the control of HCV spread throughout the developing world.

Impact of Hepatitis B and Hepatitis C Virus Infections in a Hematology-Oncology Unit at a Children's Hospital in Nicaragua, 1997 to 1999

“…Commercial reagent kits from Abbott Laboratories (North Chicago, Ill.) were used to detect HBsAg (Auszyme), total anti-HBc (Corezyme), immunoglobulin M, anti-HBc (Corezyme-M), HBeAg, anti-HBe (HBe [recombinant DNA] EIA), and anti-HBs (Abzyme); in-house reagents based on similar immunoassay principles were prepared by the Louisiana State University (LSU) International Center for Medical Research and Training (ICMRT) (2). For the screening of anti-HCV second-and third-generation commercial EIA kits (Ortho Diagnostic Systems, Raritan, N.J.) were used; EIA-ICMRT in-house (22) , commercial recombinant immunoblot assay (Ortho Diagnostic Systems) and LIA Tek III (Organon Teknika, Boxtel, The Netherlands) kits were used for confirmation of anti-HCV. Any sample being initially reactive for HBV markers by the commercial or the LSU ICMRT assays was retested to correlate the reactivity with that from a second assay.…”

“…The risk of infection with both hepatitis B virus (HBV) and hepatitis C virus (HCV) is well documented in children with hematological disorders, and prevalence rates as high as 50% in leukemia and lymphoma patients have been reported (4,21,22,26). Many of these children receive multiple transfusions of different blood components, and this could be a potential risk factor for acquiring such infections.…”

Hepatitis C virus seroprevalence in the general female population from 8 countries

“…Plasma samples were shipped to the Viral Oncology Section laboratory at Frederick National Laboratory for Cancer Research and tested for antibodies against HCV relying on a screening assay and a confirmatory test, both based on a previously reported assay (6). …”