DEVELOPING DIAGNOSTIC ASSAYS FOR BABOON CYTOMEGALOVIRUS

开发狒狒巨细胞病毒的诊断方法

• 批准号: 6394692
• 负责人: EARL BLEWETT
• 金额: $ 9.54万
• 依托单位: OSU CENTER FOR HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6394692
• 关键词: Herpesviridae disease; antiserum; baboons; communicable disease diagnosis; cytomegalovirus; diagnosis design /evaluation; diagnostic tests; enzyme linked immunosorbent assay; gene expression; molecular cloning; peptide library; virus antigen; virus classification; virus genetics; virus protein

Baboons represent an invaluable source of xenogeneic organ donors for human patients, needed to relieve the great lack of available human donor organs. However, the risk of transmitting or allowing establishment of a zoonotic virus in humans is a real, and frightening consideration. Baboons are host to a number of known viruses. This study will focus on cytomegalovirus since this is the most commonly transmitted virus in human transplantation procedures and is a major cause of many problems in immunosuppressed transplant recipients. This project will identify the indigenous CMV of baboons (BaCMV) and develop sensitive diagnostic assays. These tests will be capable of detecting BaCMV-infected potential organ donors, and useful in the screening of baboons for the establishment of specific pathogen free breeding colonies. Reagents developed in this project will also allow detection (and differentiation from HCMV) of BaCMV infection in human transplant recipients. The first goal in this project is the isolation of an indigenous CMV from baboons. This virus isolate (s) will be preliminarily characterized and used in a first generation diagnostic ELISA to detect BaCMV. This assay will be more sensitive than current detection tests since it will use BaCMV infected cells as a target antigen rather than HCMV infected cells. The most immunoreactive BaCMV proteins, as produced in a genomic X-virus expression library will be identified using CMV-positive baboon sera, cloned and sequenced. These proteins will be produced in bacterial expression systems and used in a series of second generation diagnostic assays. These strong target antigens will make the detection tests even more sensitive. In addition, the conserved CMV genes for glycoprotein B and the major immediate early protein will be cloned, sequenced and this data used to develop PCR assays to allow detection and unequivocal identification of the baboon virus in human transplant recipients.

狒狒是异种器官捐献者的宝贵来源 对于人类患者来说，需要缓解现有可用药物的严重缺乏 人类捐赠器官。然而，传输或允许的风险 人畜共患病毒在人类中的建立是真实而可怕的 考虑。狒狒是许多已知病毒的宿主。这 研究将集中于巨细胞病毒，因为这是最常见的 人类移植过程中传播的病毒是主要的 免疫抑制的移植受者出现许多问题的原因。 该项目将识别狒狒的本土巨细胞病毒（BaCMV） 并开发灵敏的诊断分析方法。这些测试将能够 检测感染 BaCMV 的潜在器官捐献者，并可用于 筛选狒狒以建立无特定病原体的环境 繁殖群落。该项目开发的试剂也将允许 BaCMV 感染的检测（以及与 HCMV 的区分） 人类移植受者。 该项目的第一个目标是 从狒狒中分离出本土巨细胞病毒。该病毒分离株 将在第一代中进行初步表征和使用 诊断 ELISA 检测 BaCMV。该检测将更加灵敏 比目前的检测测试要好，因为它将使用 BaCMV 感染细胞作为 目标抗原而不是HCMV感染的细胞。最 免疫反应性 BaCMV 蛋白，由基因组 X 病毒产生 表达文库将使用 CMV 阳性狒狒血清进行鉴定， 克隆并测序。这些蛋白质将在细菌中产生 表达系统并用于一系列第二代诊断 化验。这些强大的目标抗原将使检测测试更加顺利 更敏感。此外，糖蛋白的保守CMV基因 B 和主要的立即早期蛋白将被克隆、测序 这些数据用于开发 PCR 检测方法以进行检测和 人类移植中狒狒病毒的明确鉴定 收件人。