Parvovirus B19 genotypes 1 and 2 detection with real-time polymerase chain reaction assays

Vox Sang. 2007 Oct;93(3):208-15. doi: 10.1111/j.1423-0410.2007.00957.x.

Abstract

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.

Publication types

  • Evaluation Study

MeSH terms

  • Blood Donors
  • Genotype
  • Humans
  • Mass Screening
  • Molecular Sequence Data
  • Netherlands
  • Nucleic Acid Amplification Techniques / methods
  • Parvoviridae Infections / genetics*
  • Parvovirus B19, Human / classification
  • Parvovirus B19, Human / genetics*
  • Phylogeny
  • Polymerase Chain Reaction / methods*
  • Viral Load

Associated data

  • GENBANK/EF151136
  • GENBANK/EF151137
  • GENBANK/EF151138
  • GENBANK/EF216868