Virus and Infection

A B19V viremic serum sample, identified in our laboratory in the course of routine diagnostic analysis and available for research purposes according to Italian privacy law, was used as source of virus for the infection experiments. The viremic serum contained 10¹² B19V (genotype 1) genome copies (geq)/mL, as determined by quantitative PCR analysis, and resulted negative by routine diagnostic assays to other viruses [17].

For infection of EPCs, cells at the density of 10⁷ cells/mL were incubated in the presence of 0.1x volume of viremic serum, appropriately diluted in PBS in order to obtain a multiplicity of infection (moi) of 10³ geq/cell. Following adsorption for 2 h at 37°C, the inoculum virus was washed and the cells were incubated at 37°C in complete medium at an initial density of 5x10⁵ cells/ml, for a time course of infection.