METHODS FOR RAPID DETECTION OF CYTOMEGALOVIRUS IN BLOOD SAMPLES

巨细胞病毒(CMV)是中国人发病和死亡的主要原因。 免疫受损的宿主，尤其是在那些经历了 实体器官或骨髓移植。一个简单而敏感的人 早期检测巨细胞病毒血液感染的方法将是有帮助的 用于这些患者的医疗管理。已经做出了努力 通过检测血液中是否存在巨细胞病毒抗原 贝瓶培养和培养的巨细胞病毒pp72即刻早期抗原 CMV中性粒细胞中pp65下基质结构蛋白 用直接免疫荧光单抗检测白细胞 技术。比较敏感度、特异度、速度和 易于确定CMV抗原血症表明直接检测 对pp65抗原的检测更敏感、更快速，而且 巨细胞病毒病的早期预测因子。荧光素的直接比较- 辣根过氧化物酶标记的单抗标记 对pp65抗原的检测显示出基本相同的敏感性。 因此，我们目前使用的是免疫过氧化物酶方法 因为它的性能更快，不需要阅读 免疫荧光涂片。进一步的研究将比较 贝壳瓶培养及聚合酶链法检测抗原血症 以CMV即刻早期基因片段为引子的反应分析 承认。 努力提高贝壳小瓶化验的灵敏度 检测CMV和其他病毒的工作仍在继续。细胞经前处理 据报道，使用碘-脱氧尿嘧啶核苷可以增强复制 体外对CMV的抑制作用。我们将研究添加不同类型的 碘脱氧尿嘧啶核苷以及其他胸苷的浓度 我们的贝壳小瓶细胞培养系，以确定它们是否 提高贝壳小瓶培养检测CMV的敏感性 临床标本。

Cytomegalovirus (CMV) is a major cause of morbidity and mortality in the immunocompromised host, especially in those having undergone solid organ or bone marrow transplants. A simple and sensitive method for early detection of CMV blood infection would be helpful for the medical management of these patients. Efforts have been made to determine the presence of CMV antigen in blood by detecting both the pp72 immediate early antigen of CMV in shell vial cultures and the pp65 lower matrix structural protein of CMV in polymorphonuclear leukocytes by using the direct immunofluorescent monoclonal antibody technique. Comparisons of the sensitivity, specificity, speed, and ease of determining CMV antigenemia indicate that direct detection of the pp65 antigen is more sensitive and more rapid, as well as an earlier predictor of CMV disease. Direct comparison of fluorescein- labeled with horseradish peroxidase-labeled monoclonal antibodies for detecting pp65 antigen have shown essentially equal sensitivity. As a result, we are currently using the immunoperoxidase method because its performance is faster and is not needed to read immunofluorescent smears. Further studies will compare results of shell vial culture and the antigenemia assay with a polymerase chain reaction assay use the immediate early gene region of CMV for primer recognition. Efforts to increase the sensitivity of the shell vial assay in detecting CMV, as well as other viruses, continue. Cells pretreated with iodo-deoxyuridine have been reported to enhance the replication of CMV in vitro. We will examine the effect of adding various concentrations of iodo-deoxyuridine, as well as other thymidine analogues, to our shell vial cell culture lines to determine if they enhance the sensitivity of the shell vial culture to detect CMV in clinical specimens.