| Real-time polymerase chain reaction detection of parvovirus B19 DNA in blood donations using a commercial and an in-house assay. | |
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MedLine Citation:
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PMID: 21175647 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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BACKGROUND: European regulations require testing of manufacturing plasma for parvovirus B19 (B19) DNA to limit the load of this virus to a maximum acceptable level of 10 IU/µL. To meet this requirement, most manufacturers introduced a test algorithm to identify and eliminate high-load donations before making large manufacturing pools of plasma units. Sanquin screens all donations using a commercial assay from Roche and an in-house assay. STUDY DESIGN AND METHODS: Between 2006 and 2009, 6.2 million donations were screened using two different polymerase chain reaction (PCR) assays targeting B19 DNA. Donations with B19 DNA loads of greater than 1 × 10(6) IU/mL showing significant differences in viral load between the two assays were further analyzed by sequencing analysis. RESULTS: A total of 396 donations with B19 DNA loads of greater than 1 × 10(6) IU/mL were identified. Fifteen samples (3.8%) had discordant test results; 10 samples (2.5%) were underquantified by the Roche assay, two samples (0.5%) were underquantified by the in-house assay, and three samples (0.8%) were not detected by the Roche assay. Sequencing analysis revealed mismatches in primer and probe-binding regions. Phylogenetic analysis showed that 12 samples were B19 Genotype 1. The three samples not detected by the Roche assay were B19 Genotype 2. CONCLUSION: This study shows that 3.8% of the viremic B19 DNA-positive donations are not quantified correctly by the Roche or in-house B19 DNA assays. B19 Genotype 1 isolates showing incorrect test results are more common than B19 Genotype 2 or 3 isolates. Newly designed B19 PCR assays for blood screening should preferably have multiplexed formats targeting multiple regions of the B19 genome. |
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Authors:
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M H G M Koppelman; P van Swieten; H T M Cuijpers |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2010-12-22 |
Journal Detail:
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Title: Transfusion Volume: - ISSN: 1537-2995 ISO Abbreviation: - Publication Date: 2010 Dec |
Date Detail:
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Created Date: 2010-12-23 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0417360 Medline TA: Transfusion Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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© 2010 American Association of Blood Banks. |
Affiliation:
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From Sanquin Diagnostics, National Screening Laboratory of Sanquin (NSS) and Viral Serology, Amsterdam, the Netherlands. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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