Distribution of parvovirus B19 DNA in blood compartments and persistence of virus in blood donors.

Abstract

Because the receptor for parvovirus B19 (B19V) is on red blood cells (RBCs), we investigated B19V distribution in blood by in vitro spiking experiments and evaluated viral compartmentalization and persistence in natural infection. Two whole blood (WB) protocols (ultracentrifugation and a rapid RBC lysis and removal protocol) were evaluated using quantitative real-time polymerase chain reaction. WB was spiked with known concentrations of B19V and recovery in various blood fractions was determined. The rapid RBC lysis and removal protocol was then used to compare B19V concentrations in 104 paired WB and plasma samples collected longitudinally from 43 B19V-infected donors with frozen specimens in the REDS Allogeneic Donor and Recipient Repository (RADAR). In B19V spiking experiments, approximately one-third of viral DNA was recovered in plasma and two-thirds was loosely bound to RBCs. In the immunoglobulin (Ig)M-positive stage of infection in blood donors when plasma B19V DNA concentrations were greater than 100 IU/mL, median DNA concentrations were approximately 30-fold higher in WB than in plasma. In contrast, when IgM was absent and when the B19V DNA concentration was lower, the median WB-to-plasma ratio was approximately 1. Analysis of longitudinal samples demonstrated persistent detection of B19V in WB but declining ratios of WB to plasma B19V with declining plasma viral load levels and loss of IgM reactivity. The WB-to-plasma B19V DNA ratio varies by stage of infection, with 30-fold higher concentrations of B19V DNA in WB relative to plasma during the IgM-positive stage of infection followed by comparable levels during persistent infection when only IgG is present. Further study is required to determine if this is related to the presence of circulating DNA-positive RBCs derived from B19V-infected erythroblasts, B19V-specific IgM-mediated binding of virus to cells, or other factors.