title: Follow-up of six blood donors highlights the complementary role and limitations of hepatitis C virus antibody and nucleic acid amplification tests.
abstract: OBJECTIVE
  The purpose of this study was to analyse the follow-up results for six blood donors who screened positive for hepatitis C virus (HCV) by nucleic acid amplification technology (NAT) but were non-reactive in the primary antibody immunoassay (HCV NAT yield).
  
  
  METHODS
  Volunteer blood donations were screened, in parallel, for antibodies to hepatitis C virus (anti-HCV) and for human immunodeficiency virus (HIV)/HCV RNA using the Abbott PRISM HCV Chemiluminescent immunoassay (ChLIA) and the Chiron Procleix HIV-1/HCV RNA assays, respectively. NAT yield donor samples were further tested using supplemental assays, including an alternate HCV antibody enzyme immunoassay (EIA) (Abbott Murex anti-HCV Version 4), an immunoblot (Ortho RIBA-3 or Genelabs Diagnostics HCV Blot 3.0) and two alternative HCV NAT assays [Roche HCV Amplicor and an assembled HCV polymerase chain reaction (PCR)]. Five of the six donors were available for follow-up testing.
  
  
  RESULTS
  The six NAT yield donations were identified as constituents of 24-member minipools among 2,212,695 donations screened over the 28-month study period. All samples were positive when tested, undiluted, using the Roche Amplicor and assembled reverse transcription-polymerase chain reaction (RT-PCR) alternate NAT assays. One of the donors, subsequent to seroconversion, showed RNA levels that fluctuated above and below the limit of detection of the NAT screening assay. Three of the six were reactive on the secondary EIA and showed reactivity to the core c22(p) antigen by immunoblot at the index donation. Two others subsequently became reactive in the ChLIA prior to the EIA, showing reactivity against c100 and/or c33c antigens by immunoblot. The remaining donor became reactive in the ChLIA and EIA at the same time, showing RIBA reactivity against all of the following three peptides: c100; c33c; and c22(p).
  
  
  CONCLUSIONS
  This study demonstrated that at least five of six HCV NAT yield donors were in the pre- or early antibody seroconversion phase of infection. The observation that one yield donor demonstrated HCV RNA that fluctuated above and below the limit of detection of the primary NAT-screening assay supports the maintenance of both NAT and antibody screening for HCV. Follow-up testing of suspected yield donors revealed that the primary and alternate anti-HCV immunoassays had different performance characteristics, depending on the specificity of the donor's early anti-HCV response.
authors.affiliation: Australian Red Cross Blood Service, Brisbane, Qld., Australia. chyland@arcbs.redcross.org.au
authors.forename: C
authors.initials: C
authors.lastname: Hyland
authors.forename: C R
authors.initials: CR
authors.lastname: Seed
authors.forename: P
authors.initials: P
authors.lastname: Kiely
authors.forename: S
authors.initials: S
authors.lastname: Parker
authors.forename: N
authors.initials: N
authors.lastname: Cowley
authors.forename: W
authors.initials: W
authors.lastname: Bolton
mesh_terms: D000914:Antibodies, Viral / Q000097:blood; D001782:Blood Donors; D005188:False Negative Reactions; D005500:Follow-Up Studies; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis* / Q000635:transmission; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000592:standards*; D012367:RNA, Viral / Q000097:blood; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000592:standards*; D013997:Time Factors; D019562:Viral Load
chemical_list: D000914:Antibodies, Viral; D012367:RNA, Viral