Development of Rapid Diagnostic Tests for Hantaviruses

描述(申请人提供)：本研究方案的目标是(1)开发一种快速且易用的核酸诊断试验，以检测感染个体和携带者啮齿动物的血液、血浆或血清中的汉坦病毒；(2)开发一种快速诊断试验，以检测感染个体的血液、血浆或血清中的汉坦病毒抗体；(3)开发一种汉坦病毒核酸鉴别试验；(4)开发一种基于TaqMan的定量检测方法，以估计病毒感染样本中的汉坦病毒载量；(5)建立检测汉坦病毒心肺综合征(HCPS)患者外周血单个核细胞(PBMC)中新诺布雷病毒(SNV)抗原的流式细胞仪；(6)建立全纯合子鹿小鼠近交系；(7)建立鹿小鼠对汉坦病毒感染的细胞免疫应答的分析工具。汉坦病毒核酸和抗体诊断测试应能在一小时内得出结果，并将有助于及早发现和跟进汉坦病毒感染。汉坦病毒核酸鉴别分析的可获得性将使人们能够识别涉及汉坦病毒暴发的特定病毒分离株及其可能的来源。方法准确定量汉坦病毒在人类患者或动物模型中的载量，以及基于流式细胞术的SNV检测的可用性，近交系纯合鹿小鼠，以及分析鹿小鼠对汉坦病毒的细胞免疫反应的方法，将有助于更好地了解、预防和治疗汉坦病毒感染。这项研究涉及CHIRON公司和新墨西哥大学医学院的合作努力。这两个组织之间的前一次合作始于1994年，但在1997年终止，当时CHIRON公司撤回了对汉坦病毒诊断研究的资助，因为当时没有战略上的契合。这项任务中建议的研究产生的信息对于快速确定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.