Anti-HCV confirmatory testing of voluntary blood donors: comparison of the sensitivity of two immunoblot assays.
BACKGROUND
In this study, the sensitivity of two commercially available anti-HCV immunoblot assays (HCV Western blot (Wellcozyme] and RIBA 3.0 SIA [RIBA-3, Chiron]) was compared in a voluntary blood donor population.


METHODS
Four groups of donor samples were retrospectively tested in this study. Groups 1 and 2 were donor samples that gave positive or indeterminate band patterns, respectively, when originally tested on the HCV Western blot between 1994 and 1998. These samples were tested on the RIBA 3.0. Donor samples in Groups 3 and 4 were originally tested on RIBA-3 during 1998 and 1999 and gave positive or indeterminate blot results, respectively. In this study these two groups were tested on the HCV Western blot. Samples with discrepant results on the two immunoblot assays were selected for genotyping or serotyping.


RESULTS
The two immunoblots showed similar sensitivity to the core and NS5 proteins. However, of 35 samples positive on Western blot or RIBA-3, the Western blot failed to detect NS4 in 14 samples compared with only 5 for RIBA-3. As well, the Western blot failed to detect NS3 in 6 samples compared to 2 for RIBA-3. Five (27.8%) of 18 samples that were Western blot indeterminate due to core reactivity showed an additional NS3 band on RIBA-3. Of the samples with additional NS3 and/or NS4 reactivity on RIBA-3 that were genotyped or serotyped, all were HCV type 3.


CONCLUSIONS
Western blot and RIBA-3 showed similar sensitivity to the HCV core and NS5 proteins. However, RIBA-3 showed greater sensitivity to both NS3 and NS4 compared to the Western blot. The reduced sensitivity of the Western blot to the NS3 and NS4 proteins was observed with HCV type 3 samples.
Kiely
Philip R
PR
Virus Serology Unit, Australian Red Cross Blood Service-Victoria, South Melbourne. pkiely@arcbs.redcross.org.au
Eliades
Leo A
LA
Kebede
Mekonnen
M
Stephenson
Matthew D
MD
Jardine
Darren K
DK
D001782:Blood Donors; D015153:Blotting, Western; D016174:Hepacivirus / Q000302:isolation & purification*; D006801:Humans; D015151:Immunoblotting / Q000379:methods* / Q000592:standards*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D017361:Viral Nonstructural Proteins / Q000032:analysis; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; D012324:RNA-Dependent RNA Polymerase
D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C093542:NS4 protein, hepatitis C virus; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase