| Detection of residual donor leucocytes in leucoreduced red blood cell components using a fluorescence microplate assay. | |
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MedLine Citation:
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PMID: 12609529 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In November 1999, universal leucoreduction of blood components was introduced in the UK to minimise the risk of variant Creutzfeldt-Jakob Disease (vCJD) transmission by blood transfusion. The UK specifications for leucodepletion processes state that 99% of leucodepleted components should contain < 5 x 10(6) leucocytes/unit, within 95% confidence limits. However, this leucocyte concentration is below the detection limits of standard haematology analysers. The development of a fluorometric immunoassay to detect the residual donor leucocytes in leucoreduced blood components is described. Monoclonal antibodies to leucocyte-specific cell surface antigens, CD45 and CD15, were adsorbed to the well surface in 96-well microplates. Red blood cell samples containing low numbers of leucocytes were added to the wells and the cells of interest captured by the monoclonal antibodies. Since leucocytes are the only nucleated cells found in significant numbers in blood components they were quantified using PicoGreen, a fluorescent stain specific for dsDNA. In comparison to flow cytometry, the method currently used to detect low numbers of leucocytes, the microplate assay demonstrated excellent sensitivity (1.00) and acceptable specificity (0.81) when standard leucodepleted samples were tested. There was no significant difference between the two methods (p < or = 0.175). In conclusion, the fluorescence microplate assay represents a simple, high throughput alternative to flow cytometry for monitoring leucodepletion compliance in blood banks. |
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Authors:
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Ruth L Gilbert; Janet R Rider; J Richard Turton; Derwood H Pamphilon |
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Publication Detail:
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Type: Comparative Study; Journal Article |
Journal Detail:
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Title: Journal of immunological methods Volume: 274 ISSN: 0022-1759 ISO Abbreviation: J. Immunol. Methods Publication Date: 2003 Mar |
Date Detail:
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Created Date: 2003-02-28 Completed Date: 2003-04-09 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 1305440 Medline TA: J Immunol Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 17-25 Citation Subset: IM |
Affiliation:
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Bristol Institute for Transfusion Sciences, National Blood Service-Bristol Centre, Southmead Road, Bristol BS10 5ND, UK. rgilbert@phls.org.uk |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Blood Banks
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standards Blood Component Removal Blood Donors* Buffers Flow Cytometry Fluorescence Fluorescent Dyes Fluorometry / methods* Humans Immunoassay / methods* Leukocyte Count / methods* Organic Chemicals Reproducibility of Results Sensitivity and Specificity Time Factors |
| Chemical | |
Reg. No./Substance:
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0/Buffers; 0/Fluorescent Dyes; 0/Organic Chemicals; 0/PicoGreen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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