Development of an automated IVD for the ultra-sensitive, direct, molecular detection of Borrelia for early Lyme Disease

项目摘要 莱姆病（Lyme disease，LD）是由蜱传疏螺旋体（Borrelia）引起的最常见的病媒传染病 在美国，每年有30万新病例。如果早期诊断并使用适当的抗生素治疗， LD的结果通常很好，但治疗延迟会导致关节炎、心脏炎或神经疏螺旋体病。 早期LD最明显的表现是游走性红斑（EM）;然而，EM通常难以诊断。 与过敏反应不同，30%的感染者从未出现皮疹。非特异性 LD的症状与其他疾病相似，只能通过实验室测试来确定。电流 实验室测试依赖于血清学方法，其在诊断早期LD时无效，并且不能区分 活动性感染和既往感染之间的联系用于直接检测血液中疏螺旋体的分子诊断试验 已经证明敏感性差，因此没有获得牵引力（或FDA许可）。最终结果是 目前没有任何检测方法能够可靠地检测早期LD。 为了解决这一未得到满足的需求，改善检测LD，HelixBind开发了RaPID/LD -一个超， 专为直接检测全血中的疏螺旋体而设计的灵敏检测。RaPID/LD显示 检测限（LoD）远低于单个细胞/ml人血，灵敏度比 现有的分子诊断学。RaPID/LD仅检测活动性感染，并包含广泛的 莱姆病诱导和复发热诱导疏螺旋体，包括B。miyamotoi，并提供结果， 大约2.5小时。它已通过临床标本的初步测试，并证明其具有上级 能力的本提案重点关注：（1）完成检测试剂盒开发和初始验证测试，（2） 将当前的手动检测转换为全自动检测;包括操作的一次性塑料器械 通过小型台式仪器-能够放置在任何环境中，包括床旁，以及（3） 完成对临床标本的广泛研究，以建立性能指标。 为了在这一奋进中取得成功，我们组建了一个专家团队，在自动化的发展， 诊断由世界一流的顾问提供支持，他们在病理学、传染病、临床 微生物学、莱姆病和疏螺旋体微生物学。我们将共同加强我们的能力， 提供自动化检测，最终与我们的临床研究团队合作进行盲态、多中心临床研究。 伙伴在达到我们的特定目标后，我们将开始在ISO下进行验证和确认工作。 13485质量体系，规模生产，并完成监管许可的临床试验。RaPID/LD将 代表了计划的多重面板覆盖一系列蜱传疾病（TBD）的初始测试。

PROJECT SUMMARY Lyme disease (LD), caused by the tick-borne bacteria Borrelia, is the most common vector-borne infectious disease in the US with 300,000 new cases annually. If diagnosed early and treated with appropriate antibiotics, outcomes for LD are typically excellent, but delays in treatment result in arthritis, carditis, or neuroborreliosis. The most telling manifestation of early LD is the erythema migrans (EM); however, the EM is often difficult to distinguish from an allergic reaction, and 30% of those infected never develop the rash. Displaying non-specific symptoms which mimic those of other diseases, LD can only be confirmed by laboratory tests. Current laboratory tests rely on serological methods which are ineffective at diagnosing early LD and cannot distinguish between an active and previous infection. Molecular diagnostic tests for direct detection of Borrelia from blood have demonstrated poor sensitivity and have thus not gained traction (or FDA clearance). The end result is that no test today can detect early LD with confidence. To address this unmet need for improved detection of LD, HelixBind has developed RaPID/LD – an ultra- sensitive test specifically designed for the direct detection of Borrelia from whole-blood. RaPID/LD displays a limit of detection (LoD) well below a single cell/ml of human blood, orders of magnitude more sensitive than existing molecular diagnostics. RaPID/LD will only detect active infections and incorporates a broad menu of Lyme Disease inducing and Relapsing Fever inducing Borrelia, inclusive of B. miyamotoi, and provides results in roughly 2.5-hours. It has undergone initial testing with clinical specimens and demonstrated superior capabilities. This proposal focuses on: (1) Completing assay development and initial verification testing, (2) Translating the current manual assay to a fully automated test; comprising of single-use plastic devices operated by a small, bench-top, instrument – capable of placement in any setting inclusive of the Point-of-Care, and (3) Completing an extensive study with clinical specimens to establish performance metrics. In order to succeed in this endeavor, we have assembled a team of experts in the development of automated diagnostics supported by world-class advisors with expertise in pathology, infectious diseases, clinical microbiology, Lyme Disease, and Borrelia microbiology. Together, we will build upon our capabilities and deliver an automated assay, culminating in a blinded, multi-site, clinical study in collaboration with our clinical partners. Having achieved our Specific Aims, we will begin verification and validation efforts under an ISO 13485 Quality System, scale manufacturing, and complete clinical trials for regulatory clearance. RaPID/LD will represent the initial test for a planned multiplex panel covering a range of tickborne diseases (TBDs).