[Detection of human cytomegalovirus-antigen in peripheral blood cells by using flow-cytometer].
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.
Honda
J
J
First Department of Internal Medicine, Kurume University.
Okubo
Y
Y
Arikawa
K
K
Kumagai
M
M
Kuwamoto
M
M
Oizumi
K
K
D000328:Adult; D000956:Antigens, Viral / Q000097:blood*; D003587:Cytomegalovirus / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007962:Leukocytes / Q000276:immunology*
D000956:Antigens, Viral