Honda J, Okubo Y, Arikawa K, Kumagai M, Kuwamoto M, Oizumi K
First Department of Internal Medicine, Kurume University.
Kansenshogaku Zasshi. 1996 Sep;70(9):923-30. doi: 10.11150/kansenshogakuzasshi1970.70.923.
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.
预防巨细胞病毒（CMV）相关疾病和/或尽早开始治疗在CMV感染患者的管理中至关重要。最近，我们开发了CMV抗原血症检测法：一种基于流式细胞术检测外周血白细胞中CMV抗原的快速定量方法。从健康供体、骨髓移植患者、胶原病患者和成人T细胞白血病患者中获取肝素化外周血。为了确定HCMV感染的单核细胞的表型，使用了以下藻红蛋白偶联的单克隆抗体：CD8、CD15。将这些单克隆抗体加入全血中并孵育。溶血后，细胞用4%多聚甲醛和0.3%NP - 40固定。为了确定HCMV感染的细胞，在流式细胞术分析中使用了以下单克隆抗体：针对HCMV IEA的E13（Chemicon Inc.，东洋）。作为二抗，使用异硫氰酸荧光素（FITC）偶联的山羊抗小鼠IgG。在骨髓移植患者中，CMV相关抗原在其单核细胞和多形核细胞中呈阳性。在胶原病患者中，CMV抗原在其淋巴细胞和单核细胞中呈阳性。我们的研究表明，通过FACScan检测CMV抗原阳性血白细胞是一种快速定量方法，对CMV相关疾病的诊断和监测有用。通过FACScan进行的CMV血液抗原检测将有助于尽早开始用更昔洛韦治疗CMV相关疾病或给予预防性更昔洛韦以预防该疾病。
