Development of Rapid Diagnostic Tests for Hantaviruses

描述（申请人提供）：本研究计划的目的是（1）开发一种快速、简便的核酸诊断试验，以检测感染个体和携带者啮齿动物血液、血浆或血清中的汉坦病毒，（2）开发一种快速诊断试验，以检测感染个体血液、血浆或血清中的汉坦病毒抗体，（3）开发汉坦病毒核酸鉴别试验，（4）开发基于TaqMan的定量测定以估计病毒感染样品中的汉坦病毒载量，（5）建立流式细胞术检测外周血单个核细胞（PBMC）中新诺布雷病毒（SNV）抗原的方法（6）建立完全纯合鹿鼠的近交系亚系;（7）建立分析鹿鼠对汉坦病毒感染的细胞免疫应答的工具。汉他病毒核酸及抗体诊断测试应可于一小时内得出结果，有助及早发现汉他病毒感染及跟进。汉坦病毒核酸鉴别试验的可用性将允许鉴定涉及汉坦病毒爆发的特定病毒分离株及其可能的来源。准确定量人类患者或动物模型中汉坦病毒载量的方法，沿着基于流式细胞术的SNV测定的可用性，近交纯合鹿鼠，以及解剖鹿鼠对汉坦病毒的细胞免疫应答的方法，将有助于更好地理解、预防和治疗汉坦病毒感染。这项研究涉及凯龙公司和新墨西哥州大学医学院之间的合作努力。这两个组织之间的先前合作始于1994年，但在1997年停止，当时Chiron公司撤回了对汉坦病毒诊断研究的资助，因为当时没有战略配合。这项任务中提议的研究所产生的信息对于迅速查明可能涉及汉坦病毒制剂的生物恐怖主义袭击至关重要，对于协助医疗和公共卫生组织有效应对汉坦病毒爆发和/或威胁至关重要。

DESCRIPTION (provided by applicant): The objectives of this research proposal are (1) to develop a rapid and easy-to-use nucleic acid diagnostic test to detect hantaviruses in blood, plasma, or serum of infected individuals and carrier rodents, (2) to develop a rapid diagnostic test to detect hantavirus antibodies in blood, plasma, or serum of infected individuals, (3) to develop a hantavirus nucleic acid discriminatory test, (4) to develop TaqMan-based quantitative assays to estimate hantavirus load in virus infected samples, (5) to develop a flow cytometry assay to detect Sin Nombre virus (SNV) antigen in peripheral blood mononuclear cells (PBMC) of patients with hantavirus cardiopulmonary syndrome (HCPS), (6) to develop inbred sublines of fully homozygous deer mice, and (7) to establish tools to dissect deer mouse cellular immune responses to hantavirus infection. The hantavirus nucleic acid and antibody diagnostic tests should be able to yield results in less than one hour, and will assist in the early detection and follow up of hantavirus infections. The availability of the hantavirus nucleic acid discriminatory assay will allow identification of specific viral isolates involved in hantavirus outbreaks and their possible origins. Methods to accurately quantitate hantavirus load in human patients or in animal models, along with the availability of a flow cytometry-based SNV assay, inbred homozygous deer mice, and methodologies to dissect the deer mouse cellular immune responses to hantavirus will facilitate better understanding, prevention, and treatment of hantavirus infections. This research involves collaborative efforts between Chiron Corporation and the School of Medicine, University of New Mexico. A previous collaboration between the two organizations was initiated in 1994 but discontinued in 1997 when Chiron Corporation withdrew funding for hantavirus diagnostic research, as there was no strategic fit at that time. The information generated from the studies proposed in this task will be crucial for the rapid identification of possible bioterrorism attacks involving hantavirus agents and will be vital to assist the medical and public health organizations to efficiently respond to hantavirus outbreaks and/or threats.