Diagnostic assays for Borrelia Ionestari

描述（由申请人提供）：南方蜱相关皮疹病（STARI）是一种新描述的莱姆病样感染，报告来自美国东南部和中南部，由疏螺旋体引起。我们的长期目标是了解lonestari作为人类病原体的重要性。本应用程序的目的是发展直接的显微镜，培养，分子和血清学方法，将允许诊断人类，实验动物模型和宿主中的lonestari感染。疏螺旋体与宫本氏疏螺旋体、氏疏螺旋体和回归热疏螺旋体亲缘关系最密切，它们在感染期间都会产生高密度螺旋体。感染回归热伯氏疏螺旋体也会导致强烈的体液免疫反应。我们对两只鹿小鹿的初步试验感染表明，白螺旋体也产生循环螺旋体，并导致受感染动物的血清转化。因此，提出的研究背后的基本原理是，像感染回归热伯氏疏螺旋体一样，lonestari感染可能导致可检测的螺旋体血症，允许从血液中培养生物体，在血液涂片上显示，并通过分子分析进行检测，以及强烈的抗体反应，这将允许开发免疫诊断工具。为了实现这一应用的目标，我们将追求三个具体目标：(1)改进我们的培养方法，并利用从实验感染的鹿身上收集的样本直接识别循环血液中的螺旋体；(2)建立疏螺旋体分子鉴定方法，使其能够与伯氏疏螺旋体和回归热疏螺旋体快速、可靠地区分；(3)鉴定候选抗原，这些候选抗原可能引发产生孤芽孢杆菌特异性抗体。在这项研究完成后，我们期望开发出培养、显微镜、分子和血清学技术，以特异性地识别B. lonestari感染。这项拟议的研究所产生的诊断分析将使人们能够准确地识别导致人类疾病的疏螺旋体，并最终更有力地了解疏螺旋体作为人类病原体的作用。结果应能提高对人类lonestari感染的诊断。

DESCRIPTION (provided by applicant): Southern tick-associated rash illness (STARI) is a newly described Lyme disease-like infection reported from the southeastern and south central US caused by Borrelia lonestari. Our long-range goal is to understand the importance of B. lonestari as a human pathogen. The objective of this application is to develop direct microscopic, culture, molecular, and serologic methods that will allow diagnosis of B. lonestari infection in humans, experimental animal models, and reservoir hosts. Borrelia lonestari is most closely related to B. miyamotoi, B. theileri, and the relapsing fever Borrelia, all of which produce high-density spirochetemia during infection. Infection with relapsing fever Borrelia also results in a strong humoral immune response. Our pilot trial infection of two deer fawns showed that B. lonestari also produces a circulating spirochetemia and results in seroconversion of infected animals. The rationale behind the proposed research, therefore, is that like infection with relapsing fever Borrelia spp., B. lonestari infection is likely to result in detectable spirochetemia, allowing culture of the organism from blood, visualization on blood smears, and detection by molecular assays, as well as a strong antibody response which will permit development of immunodiagnostic tools. To accomplish the objectives of this application, we will pursue three specific aims: (1) refine our methods for culturing B. lonestari and for directly identifying spirochetes in circulating blood using samples collected from experimentally infected deer; (2) develop molecular assays for identification of Borrelia lonestari that allow it to be quickly and reliably distinguished from B. burgdorferi and relapsing fever Borrelia; and (3) identify candidate antigens that may elicit production of B. lonestari-specffic antibodies. At the completion of this research, we expect to have developed culture, microscopic, molecular, and serologic techniques that will specifically identify B. lonestari infection. The diagnostic assays resulting from this proposed research will allow accurate identification of the Borrelia spp. causing disease in people and, ultimately, a more robust understanding of the role of B. lonestari as a human pathogen. Improved diagnosis of human B. lonestari infection should result.