Parvovirus B19 genotypes 1 and 2 detection with real-time polymerase chain reaction assays.
| Autor: | Koppelman MH; Sanquin, Diagnostic Services Division, Amsterdam, The Netherlands. m.koppelman@sanquin.nl, Rood IG, Fryer JF, Baylis SA, Cuypers HT |
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| Jazyk: | angličtina |
| Zdroj: | Vox sanguinis [Vox Sang] 2007 Oct; Vol. 93 (3), pp. 208-15. |
| DOI: | 10.1111/j.1423-0410.2007.00957.x |
| Abstrakt: | Background and Objectives: 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. Materials and 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. |
| Databáze: | MEDLINE |
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