Assessing the effect of removing human cytomegalovirus from blood components using quantitative polymerase chain reaction
LU Ping1；LING Bing1；FANG Feng-qin2；JI Yu-hua2
1. Shanghai Blood Center, Shanghai 200051, China; 2. Clinical Diagnostic Laboratory, Rujin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
The purpose of the current study is to assess the effect of removing human cytomegalovirus ( HCMV) by leukocyte reduction filtration ( LRF) in blood center. Peripheral blood mononuclear cells ( PBMCs) from blood samples negative for HCMV-IgG and pp65 were separated and co-cultured with HCMV-infected human fibroblasts in order to establish infection. The infected PBMCs were purified from fibroblasts by double selection using magnetic beads coated with anti-human fibroblast antibodies and anti-CD45 and then spiked into whole blood units prior to LRF. After LRF, HCMV load in whole blood and in fibroblasts was assessed using real-time quantitative polymerase chain reaction( PCR) for HCMV DNA and quantitative reverse transcriptase PCR( RT-PCR) for mRNA encoding the HCMV DNA and fibroblast-specific splice variant of prolyl-4-hydroxylase, respectively. After LRF, the rate of leukocyte depletion in whole blood units ( 200 ml) was 99. 98%. After correcting for fibroblast-associated HCMV, the mean HCMV load in whole blood was reduced from3 742 to 22. 57 copies per microliter ( 2. 50 log10 reduction) by LRF. These results suggest that LRF reduces viral load but does not completely remove HCMV from blood components.
