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 Blood, 1 February 2004, Vol. 103, No. 3, pp. 1137-1139.
Prepublished online as a Blood First Edition Paper on October 2, 2003; DOI 10.1182/blood-2003-03-0762.

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Submitted March 11, 2003
Accepted September 3, 2003

Assessment of removal of human cytomegalovirus from blood components by leucocyte depletion filters using real-time quantitative PCR

Micaela Rios Visconti, Joanne Pennington, Stephen F Garner, Jean-Pierre Allain, and Lorna M Williamson*

Division of Transfusion Medicine, University of Cambridge, Cambridge, United Kingdom; National Blood Service, Cambridge, United Kingdom

* Corresponding author; email: lorna.williamson{at}nbs.nhs.uk.

To assess removal of CMV by leucocyte depletion (LD) filters, we developed a spiking model of latent virus, using peripheral blood mononuclear cells (PBMC) infected by co-culture with CMV-infected human fibroblasts. Infected PBMC were purified by dual magnetic column selection, then spiked into whole blood units or buffy coat pools prior to LD by filtration. CMV load and fibroblast contamination were assessed using quantitative CMV DNA real-time PCR and quantitative RT-PCR of mRNA encoding the fibroblast-specific splice variant of prolyl-4-hydroxylase, respectively. After correcting for fibroblast-associated CMV, the mean CMV load was reduced in whole blood by LD from 7.42 x 102 to 1.13 copies/µl (2.8110log reduction), and from 3.8 x 102 to 4.77 copies/µl (1.910log reduction) in platelets. These results suggest that LD by filtration reduces viral burden but does not completely remove CMV from blood components.


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