Detection of human cytomegalovirus (HCMV) DNA from cell-free plasma of immunosuppressed patients by nested PCR: influence of nucleic acid extraction method

Conclusions Blood cells and plasma preparations from HCMV-seropositive healthy blood donors were all nPCR negative. Detection of HCMV DNA from PBMC and granulocytes (DNAemia) of immunosuppressed patients by nPCR did not correlate with the isolation of infectious virus from these cell populations in cell culture (viremia). However HCMV could be isolated in 60% of cases from other materials of the same patient. HCMV DNA detected in blood cells persisted for up to one year in an asymptomatically infected individual after NTX. The sensitivity of HCMV DNA detection in cell-free plasma (up to 5 fg) depended on the method used for DNA isolation. The rate of HCMV DNA detection in plasma was lower than in leukocytes. In all cases of positive plasma PCR infectious virus could be isolated from any other material of the symptomatically infected patients. Therefore HCMV DNA PCR from plasma of immunosuppressed patients seems to be a suitable and easy alternative to HCMV RT/PCR for routine diagnosis of HCMV disease.