Parvovirus B19 genotypes 1 and 2 detection with real-time polymerase chain reaction assays.
OBJECTIVE
Parvovirus B19 (B19V) DNA screening has been introduced to comply with European regulations for certain plasma products. Current commercial and some in-house B19V DNA assays fail to detect or under-quantify the recently identified genotypes 2 and 3. In this report, we describe 2-year experience with B19V DNA screening using the commercial assay from Roche (detecting only genotype 1) combined with an in-house assay (detecting genotypes 1, 2 and 3). This dual testing approach enables the identification of molecular variants of B19V.


METHODS
Between 2005 and 2007, approximately 2.6 million plasma donations were screened for B19V DNA loads exceeding 10(6) IU/ml using the Roche and the in-house real-time polymerase chain reaction assay.


RESULTS
A total of 232 plasma units were identified with B19V DNA loads above 10(6) IU/ml. Concordant results were observed for the majority of B19V positive samples; however, three of these showed discrepant results between the two assay systems. One was a B19V genotype 2 strain not detected by the Roche assay; another was a B19V genotype 1 strain with a mismatch in the 3'-end of the reverse primer and therefore under-quantified by the Roche assay; and the third one was also a B19V genotype 1 strain that gave an unusual amplification plot in the in-house assay due to a mismatch in the probe-binding site.


CONCLUSIONS
New, high viral load, B19V genotypes 2 and 3 infections are rare in blood donors tested by Sanquin. One case was found while testing 2.6 million donations. The prevalence of B19V genotype 1 variants not detected by commercial or in-house assays might be in the same range or even higher than the prevalence of B19V genotype 2 viruses, which remain undetected.
Koppelman
M H G M
MH
Sanquin, Diagnostic Services Division, Amsterdam, The Netherlands. m.koppelman@sanquin.nl
Rood
I G H
IG
Fryer
J F
JF
Baylis
S A
SA
Cuypers
H T M
HT
D001782:Blood Donors; D005838:Genotype; D006801:Humans; D008403:Mass Screening; D008969:Molecular Sequence Data; D009426:Netherlands; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D010322:Parvoviridae Infections / Q000235:genetics*; D016732:Parvovirus B19, Human / Q000145:classification / Q000235:genetics*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D019562:Viral Load