A method for the detection and confirmation of antibodies to hepatitis C virus in dried blood spots

J Virol Methods. 1997 Nov;68(2):199-205. doi: 10.1016/s0166-0934(97)00127-4.

Abstract

This study describes the development and evaluation of a cost effective test rationale for the detection of anti-HCV in dried blood spots. Samples were screened using an 'in house' IgG ELISA that incorporated the recombinant proteins c22-3, c200 and NS5. Confirmation of specific antibody to HCV was by a modification of the immunoblot RIBA 3.0. An extensive panel of well evaluated anti-HCV positive and negative samples from the UK and South Africa were used to assess the sensitivity and specificity of the two tests. One third of the anti-HCV positive samples had been typed. All anti-HCV positive samples were detected by the 'in house' screening EIA. Test/negative optical density ratios showed that more than 95% of reactive samples produced values greater than 5.0. Antibodies to HCV could be detected in a wide range of samples derived from asymptomatic and symptomatic patients and of different genotypes, with similar sensitivity. The presence of anti-HCV could be confirmed by RIBA 3.0 in samples with low reactivity but not in anti-HCV negative samples. Furthermore the immunoblot assay successfully increased specificity by screening out false reactive EIA samples that might occur in an epidemiological survey of a multi-ethnic population.

PIP: While hepatitis C virus (HCV) is a major etiological agent of post-transfusion and community acquired non-A, non-B hepatitis, little is known about the epidemiology of HCV in the UK. A cost-effective method using dried blood spots to determine anti-HCV IgG in subjects which could be used in large-scale epidemiological studies is described. Samples were screened using an in-house IgG ELISA incorporating the recombinant proteins c22-3, c200, and NS5, while specific antibody to HCV was confirmed using a modified immunoblot RIBA 3.0. A panel of well evaluated anti-HCV positive and negative samples from the UK and South Africa were used to assess the sensitivity and specificity of the 2 tests. All anti-HCV positive samples were detected by the in-house screening EIA. Test/negative optical density ratios showed that more than 95% of reactive samples produced values greater than 5.0. Antibodies to HCV could be detected in a wide range of samples derived from asymptomatic and symptomatic patients and of different genotypes, with similar sensitivity. The presence of anti-HCV could be confirmed by RIBA 3.0 in samples with low reactivity, but not in anti-HCV negative samples. The immunoblot assay increased specificity by screening out false reactive EIA samples.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antibodies, Viral / blood*
  • Female
  • HIV-1 / immunology
  • HIV-2 / immunology
  • Hepacivirus / immunology*
  • Human T-lymphotropic virus 1 / immunology
  • Humans
  • Immunoblotting / methods*
  • Immunoenzyme Techniques*
  • Immunoglobulin G / blood
  • Infant, Newborn
  • London
  • Neonatal Screening
  • Reagent Kits, Diagnostic
  • Sensitivity and Specificity
  • South Africa

Substances

  • Antibodies, Viral
  • Immunoglobulin G
  • Reagent Kits, Diagnostic