METHODS FOR RAPID DETECTION OF CYTOMEGALOVIRUS IN BLOOD SAMPLES

快速检测血样中巨细胞病毒的方法

• 批准号: 2571427
• 负责人: H D ENGLER
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2571427
• 关键词: Herpesviridae disease; blood tests; communicable disease diagnosis; cytomegalovirus; diagnosis design /evaluation; human tissue; idoxuridine; immunofluorescence technique; immunoperoxidase; microorganism culture; neutrophil; polymerase chain reaction; rapid diagnosis; virus antigen; virus load; virus protein

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.

巨细胞病毒（CMV）是一个主要的原因，发病率和死亡率， 免疫功能低下的宿主，特别是那些经历过 实体器官或骨髓移植一个简单而敏感的 一种早期发现CMV血液感染的方法将是有帮助的 对这些病人进行医疗管理。已作出努力 为了确定血液中CMV抗原的存在， 壳瓶培养物中CMV的pp 72立即早期抗原， pp 65降低CMV基质结构蛋白多态性 白细胞直接免疫荧光单克隆抗体 法比较灵敏度、特异性、速度和 确定CMV抗原血症的容易性表明直接检测 pp 65抗原的检测更敏感、更快速， CMV疾病的早期预测因子。荧光素的直接比较- 辣根过氧化物酶标记单克隆抗体标记 用于检测PP 65抗原的灵敏度基本相同。 因此，我们目前使用免疫过氧化物酶法 因为它的性能更快， 免疫荧光涂片。进一步的研究将比较 壳瓶培养和聚合酶链抗原血症测定 以CMV的即早基因区为引物， 识别. 努力提高壳小瓶测定法的灵敏度， 检测CMV以及其他病毒的工作仍在继续。预处理的细胞 与碘脱氧尿苷已经报道，以提高复制 CMV在体外我们将研究添加各种 碘脱氧尿苷以及其他胸苷的浓度 类似物，我们的壳小瓶细胞培养系，以确定他们是否 提高壳瓶培养检测CMV的敏感性， 临床标本