Prophlaxis of CMV-associated disease and/or early initiation of
therapy is important in the management of patients with CMV
infection. Recently, we developed the CMV antigenemia assay: a rapid and quantitative method based on the detection of CMC
antigens in peripheral blood leukocytes by flow cytometry. Heparinized peripheral blood was obtained from healthy donors,
bone marrow transplantation patients, patient with
collagen disease and patients with
adult T cell leukemia. To determine the phenotype of HCMV-infected mononuclear cells, the following
phycoerythrin-conjugated mAb were used: CD8, CD15. These mAds were added to the whole blood and incubated. After the
hemolysis, the cells were fixed with 4%
paraformaldehyde and 0.3%
NP-40. To determine the HCMV-infected cells, the following mAb were used in flow cytometry analysis: E13 (Chemicon Inc., Toyo) against HCMV IEA. As a secondary antibody, a
FITC-conjugated goat anti-mouse
IgG. In the bone marrow transplant patients, CMV-associated
antigen was positive in their monocytes and polymorphocytes. In the patient with
collagen disease, CMV-
antigen was positive in their lymphocytes and monocytes. Our study demonstrates that the detection of CMV
antigen-positive blood leukocytes by FACScan is a rapid and quantitative method and useful for the diagnosis and monitoring of CMV-associated CMV-associated disease. The CMV blood
antigen assay by FACScan will facilitate the initiation of early treatment with
ganciclovir of CMV-associated disease or the administration of prophylactic
ganciclovir for preventing the disease.