Evaluation of indeterminate c22-3 reactivity in volunteer blood donors.
BACKGROUND
Approximately 25 percent of blood donor sera that are repeatably reactive for hepatitis C virus (HCV) on second-generation enzyme immunoassay (EIA 2.0) are indeterminate on second-generation recombinant immunoblot assay (RIBA 2.0), and over 76 percent of these results are due to single reactivity to the HCV recombinant antigen c22-3.


METHODS
Data are presented on 46 volunteer allogeneic blood donors who were reactive on EIA2.0 and c22-3 indeterminate in RIBA 2.0. Index and follow-up samples were evaluated by using a panel of five synthetic peptide EIAs, a prototype strip immunoblot assay that uses synthetic peptides in addition to recombinant protein (RIBA 3.0), and polymerase chain reaction (PCR) for HCV RNA.


RESULTS
All 46 donations had normal alanine aminotransferase values; only 2 (4.3%) reacted for antibody to hepatitis B core antigen. With a panel of 12 synthetic peptides spanning the entire sequence of the c22-3 recombinant antigen, 33 plasmas (72%) reacted to one peptide or none, including 19 plasmas with reactivity restricted entirely to the N-terminal peptide (1-15 amino acids) of c22-3. With RIBA 3.0, 28 donations (61%) were nonreactive, including 25 that reacted with one peptide or none in EIA. Of these 25 plasmas, 18 reacted with the N-terminal sequence only. All 46 index donations were tested by PCR; the single PCR-positive unit reacted with four HCV peptides, was positive by RIBA 3.0, and reacted for antibody to hepatitis B core antigen. Twenty-six index donors were successfully recalled 3 to 7 months after their index donation. None seroconverted to positivity in RIBA 2.0, 1 was nonreactive, and 25 remained positive for c22-3 only. The restricted epitope reactivity in peptide EIA and RIBA 3.0 was maintained over time in all cases. All 26 of the follow-up samples tested negative by PCR.


CONCLUSIONS
On the basis of the restricted peptide reactivity and PCR negativity of index and follow-up samples, it is concluded that the majority of c22-3 RIBA 2.0-indeterminate results are due to nonspecific cross-reactivity to restricted (principally, N-terminal) regions of HCV core antigen.
Tobler
L H
LH
Irwin Memorial Blood Centers, University of California, San Francisco.
Busch
M P
MP
Wilber
J
J
Dinello
R
R
Quan
S
S
Polito
A
A
Kochesky
R
R
Bahl
C
C
Nelles
M
M
Lee
S R
SR
D000410:Alanine Transaminase / Q000097:blood; D000595:Amino Acid Sequence; D000941:Antigens / Q000276:immunology*; D001782:Blood Donors; D004797:Enzyme-Linked Immunosorbent Assay; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D011994:Recombinant Proteins / Q000276:immunology; D014838:Volunteers
D000941:Antigens; D012367:RNA, Viral; D011994:Recombinant Proteins; D000410:Alanine Transaminase