Broad Detection System for Tick-Borne Pathogens

描述(由申请人提供)：在美国，莱姆病和其他由壁虱传播的疾病的病例和传播越来越多。可以传播莱姆病病原体的同一只扁虱，伯氏疏螺旋体，也可以同时传播其他病原体，如巴贝斯病、无浆体和立克次体。这是公众和他们的医生非常关注的问题。这种担忧反映在对检测的更大需求上，这些检测可以提供关于单个滴虫中有什么以及可以传播什么的信息。目前对这些致病病原体的诊断检测方法通常限于单一微生物和/或不可靠。目前还没有一种检测方法可以覆盖由虱子传播的病原体的广度。在我们的第一阶段应用中，我们实现了我们的目标，概念证明，我们的方法可以检测多个微生物，甚至在一个滴答中。该系统采用了广泛的聚合酶链式反应和质谱学(聚合酶链式反应/电喷雾质谱)。该检测鉴定并形成了广泛的细菌群体，包括已知的壁虱传播细菌和黄病毒。这一成功，再加上不断增长的需求和仅针对疏螺旋体进行的搜索次数的增加，是第二阶段集中开发利用高通量PCR/ESI-MS进行壁虱传播病原体检测的基础。在第一阶段，病原体检测原理的证明是在一个96个配套板的16个孔中用16对引物完成的。在第二阶段，目标1是增加临床上有用的检测时间和效率，从而降低成本。我们将通过消除冗余和多路传输来实现8个井(聚合酶链式反应)的目标。目标2：优化并添加巴贝斯虫检测引物。目标3：建立一个可补充的从受感染的扁虱中提取核酸的资料库，用于未来对这类化验的研究以及扁虱-人类界面研究。这家银行将具有商业价值。在第二阶段完成后，我们将能够为研究人员、医生、公共卫生实体、私人和个人公民部门提供壁虱微生物诊断服务。这将是管理莱姆病和相关的壁虱传播疾病的一个重要工具，以改善医疗和公共卫生监测。它也将是对患者的皮肤、血液、关节液和脑脊液进行同样处理的基础。与公共卫生的相关性：准确识别扁虱中能够引起莱姆病和相关疾病的多种病原体的能力，将在暴露于扁虱或被扁虱叮咬的患者的管理策略中发挥重要作用。这将有助于降低患者和公共卫生系统不断上升的成本。

DESCRIPTION (provided by applicant): There is an increasing number of cases and spread of Lyme and other tick-borne diseases in the United States. The same tick that can transmit the agent of Lyme disease, Borrelia burgdorferi, can simultaneously transmit other pathogens such as Babesia, Anaplasma, and Rickettsia. This is of great concern to the public and their physicians. This concern is reflected in a greater demand for tests that can provide information as to what is in an individual tick and what can be transmitted. Current diagnostic testing methods for these disease causing pathogens are generally limited to a single microbe and/or unreliable. No test exists that can cover the breadth of pathogens transmitted by ticks. In our Phase I application, we achieved our aim, the proof of concept, that our methodology could detect multiple microbes and even in a single tick. That system employs broad-range PCR and mass spectroscopy (PCR/ESI-MS). The assay identified and speciated broad groups of bacteria, including known tick- borne bacteria and flaviviruses. The success of this, coupled with the increasing demand and increasing number of searches performed for Borrelia alone, is the basis for Phase 2 to concentrate on the development of a tick-borne pathogen detection assay using the high throughput PCR/ESI-MS. In Phase 1, the proof of principle of detection of pathogens was accomplished with 16 primer pairs in 16 wells of a 96 well kitted plate. In Phase 2, Goal 1 of this is to increase clinically useful throughput time and efficiency of the assay and consequently reduce costs. We will have a target goal of 8 wells (PCR reactions) by eliminating redundancy and multiplexing. Goal 2: Optimize and add to these, Babesia detection primers. Goal 3: Create a replenishable bank of nucleic acid extraction from infected ticks for future investigation of this type of assay as well as tick-human interface research. This bank will be of commercial value. Upon the completion of Phase2, we will be in a position to provide a tick-microbial diagnostic service for researchers, physicians, public health entities, private and individual citizen sectors. This will be an important tool in management of Lyme and related tick-borne diseases for improved medical treatment and public health surveillance. It will also be the foundation for doing the same in patient skin, blood, joint fluid, and cerebrospinal fluid. PUBLIC HEALTH RELEVANCE: The ability to accurately identify multiple pathogens in the ticks capable of causing Lyme disease and related disorders will be important in management strategy of patients exposed or bitten by ticks. It will help reduce rising costs to the patient and the public health system.