Diagnostic assays for Borrelia Ionestari

描述(由申请人提供)：南方硬虱相关皮疹疾病(STARI)是一种新发现的莱姆病样感染，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孤独杆菌作为人类病原体的重要性。这项应用的目标是开发直接的显微镜、培养、分子和血清学方法，以便能够诊断人类、实验动物模型和水库宿主中的孤独芽孢杆菌感染。与米氏疏螺旋体、泰勒氏疏螺旋体和回归热疏螺旋体关系最密切的是疏螺旋体，所有这些都会在感染期间产生高密度螺旋体血症。感染回归热疏螺旋体也会引起强烈的体液免疫反应。我们对两只小鹿的初步感染试验表明，长圆线虫也会产生循环螺旋体血症，并导致受感染动物的血清转换。因此，这项拟议研究背后的基本原理是，就像感染回归热疏螺旋体一样，孤独芽孢杆菌感染可能会导致可检测到的螺旋体血症，允许从血液中培养有机体，在血液涂片上进行可视化，通过分子分析进行检测，以及强大的抗体反应，这将使免疫诊断工具的开发成为可能。为了实现这一应用程序的目标，我们将追求三个具体目标：(1)改进我们的方法，以培养孤独螺旋体，并使用从实验感染的鹿身上收集的样本直接鉴定循环血液中的螺旋体；(2)建立鉴定孤独螺旋体的分子检测方法，使其能够快速可靠地与伯氏疏螺旋体和复发性热疏螺旋体区分开来；以及(3)鉴定可诱导产生孤独螺旋体特异性抗体的候选抗原。在这项研究完成后，我们预计将开发出培养、显微镜、分子和血清学技术，专门鉴定孤独芽胞杆菌感染。这项拟议研究产生的诊断化验将使疏螺旋体的准确鉴定成为可能。在人类中引发疾病，并最终对孤独杆菌作为人类病原体的作用有了更深入的了解。人类孤独芽孢杆菌感染的诊断应该会有所改善。

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.