title: Validation of a strategy for HCV antibody testing with two enzyme immunoassays in a routine clinical laboratory.
abstract: BACKGROUND
  Centers for Disease Control (CDC) guidelines require confirmation of hepatitis C virus (HCV) screening-test-positive sera with a low signal/cut-off (S/CO) ratio by recombinant immunoblot or PCR. The UK Health Protection Agency has suggested that a second enzyme immunoassay (EIA) could be used as an alternative for confirmation in non-immunocompromised patients.
  
  
  OBJECTIVE
  To evaluate the UK HPA approach in 17,936 consecutive in-house sera submitted for HCV testing.
  
  
  METHODS
  AxSYM-positive sera (S/CO> or =1.0) were tested with Monolisa Plus. AxSYM-positive sera of patients that were confirmed PCR-positive were considered HCV+. All other AxSYM-positive sera were confirmed with immunoblot according to CDC guidelines.
  
  
  RESULTS
  17,299 sera were negative with AxSYM. Of the 637 AxSYM-positive sera, 384 were from patients confirmed as PCR-positive. Of other 250 sera, 120 were negative with immunoblot, 103 were positive and 30 were indeterminate. All 30 immunoblot-indeterminate sera were PCR-negative. Two patients were Monolisa Plus+ and immunoblot- and PCR-. One patient was known as immunoblot-, while the other patient was diagnosed with non-A non-B hepatitis in 1980s. Nine sera from HCV-positive patients were Monolisa Plus-. Two PCR- sera were from immunocompetent patients who were PCR- for > or =8 years and six PCR- sera and one PCR+ serum were from immunocompromised patients. Sensitivity and specificity of confirmation with Monolisa Plus were 98.15% and 98.33% and the positive and negative predictive values were 99.58% and 92.91% in AxSYM-positive sera (excluding immunoblot-indeterminate/PCR-negative sera). If immunocompromised patients that were false-negative were excluded, sensitivity was 99.58%.
  
  
  CONCLUSIONS
  Monolisa Plus can be used as an alternative to immunoblot for the confirmation of AxSYM-positive sera.
authors.affiliation: Department of Laboratory Medicine, University of Leuven, Leuven, Belgium. pieter.vermeersch@uz.kuleuven.be
authors.forename: Pieter
authors.initials: P
authors.lastname: Vermeersch
authors.forename: Marc
authors.initials: M
authors.lastname: Van Ranst
authors.forename: Katrien
authors.initials: K
authors.lastname: Lagrou
mesh_terms: D016174:Hepacivirus / Q000276:immunology; D006526:Hepatitis C / Q000175:diagnosis* / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000097:blood*; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007124:Immunoenzyme Techniques / Q000379:methods; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000235:genetics; D012680:Sensitivity and Specificity; D006113:United Kingdom
chemical_list: D018937:Hepatitis C Antibodies; D012367:RNA, Viral