Development of an EBV Real-time PCR Assay and Application to Two Epidemiological Studies

A real-time quantitative FOR assay capable of measuring EBV DNA load was developed, validated and applied to test hypotheses relevant to EBV associated disease and pathogenesis. EBV appears to be a major cofactor that predisposes iatrogenically and naturally immunocompromised individuals, such as HIV-infected subjects, transplant patients and others, to develop various lymphoproliferative disorders. Recent evidence suggests that these patients experience elevated EBV levels in PBMC, blood and saliva. To determine whether EBV loads are elevated in PBMC of HIV infected subjects compared to immunocompetent controls and whether EBV loads are higher in EBV related AIDS lymphoma subjects compared to HIV infected subjects without cancers, we studied a subset of 459 HIV-1 seropositive patients with and without cancers and 61 randomly selected blood donors. Our results showed that EBV load was significantly increased when subjects were HIV positive and even higher if they had cancers, as compared to immunocompetent controls. Additional results from a Ugandan sickle cell disease study of 600 children (and their mothers), were in agreement with these findings irrespective of age or gender of the subjects. Uganda is an area endemic for BL and KSHV.