Rare detection of hepatitis B and hepatitis C virus genomes by polymerase chain reaction in seronegative donors with elevated alanine aminotransferase.
BACKGROUND
Since screening for antibody to hepatitis C virus (HCV) was introduced in 1990, posttransfusion hepatitis has been reduced to nearly background levels. This has led to reconsideration of the value of testing donated blood for elevated alanine aminotransferase (ALT). The contribution of ALT testing in detecting seronegative infection was evaluated by the performance of polymerase chain reaction (PCR) for hepatitis B virus (HBV) or HCV in plasma from ALT-elevated blood units.


METHODS
Testing was performed on 375 units of plasma, derived from an equivalent of 47,500 blood donations, with a highly sensitive hemi-nested PCR procedure. Using a triplet of primers directed at the conserved regions of HBV DNA and 5'-noncoding regions of HCV RNA, the hemi-nested PCR assay can reliably amplify 10 viral molecules to levels detectable in ethidium bromide-stained agarose gels. Pools of plasma from groups of four donors were screened with hemi-nested PCR. For any reactive pools, the plasma from individual donors was retested twice on different aliquots.


RESULTS
Two of 375 units, both with midrange ALT elevation, were repeatedly reactive in hemi-nested PCR (one each for HBV DNA and HCV RNA). However, samples from the two suspect donors tested 9 and 5 months later revealed no seroconversion, elevated ALT, or viral genomes in hemi-nested PCR.


CONCLUSIONS
The lack of confirmed HBV or HCV infection in this study representing an estimated 47,500 voluntary blood donations suggests that routine ALT testing for further prevention of posttransfusion hepatitis after exclusion of HBV- and/or HCV-seropositive blood may be superfluous.
Sankary
T M
TM
Department of Laboratory Medicine, University of California, San Francisco.
Yang
G
G
Romeo
J M
JM
Ulrich
P P
PP
Busch
M P
MP
Rawal
B D
BD
Vyas
G N
GN
D000410:Alanine Transaminase / Q000097:blood*; D001483:Base Sequence; D004279:DNA, Viral / Q000097:blood; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification; D006509:Hepatitis B / Q000175:diagnosis*; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification; D006526:Hepatitis C / Q000175:diagnosis*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012698:Serologic Tests
D004279:DNA, Viral; D000410:Alanine Transaminase