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

项目总结 莱姆病(LD)是最常见的媒介传播传染病，由壁虱传播的疏螺旋体细菌引起。 这种疾病在美国每年有30万新病例。如果及早诊断并使用适当的抗生素治疗， LD的结果通常很好，但延误治疗会导致关节炎、心脏炎或神经疏螺旋体病。 早期LD最明显的表现是移行性红斑(EM)；然而，EM通常很难 与过敏反应不同的是，30%的感染者从未出现皮疹。显示非特定 LD的症状与其他疾病相似，只有通过实验室测试才能确认。当前 实验室检测依赖于血清学方法，这些方法在诊断早期LD方面无效，并且无法区分 活动性感染和前次感染之间的关系。直接从血液中检测疏螺旋体的分子诊断试验 表现出较差的敏感性，因此没有获得牵引力(或FDA批准)。最终的结果是 如今，没有一项测试能够可靠地检测出早期的LD。 为了解决这一未得到满足的改进LD检测的需求，HelixBind开发了RAPID/LD-一种超 专为直接从全血中检测疏螺旋体而设计的敏感试验。RAPID/LD显示 检出限(LOD)远低于单细胞/毫升人体血液，比 现有的分子诊断学。RAPID/LD将只检测活动性感染，并包含一个广泛的菜单 莱姆病诱发和复发性发热诱导疏螺旋体，包括宫本杆菌，并在 大约2.5个小时。它已经通过了临床标本的初步测试，并被证明是优越的 能力。这一建议的重点是：(1)完成化验开发和初步验证测试，(2) 将目前的手工化验转变为完全自动化的化验；由一次性使用的塑料设备组成 通过一个小型的台式仪器，能够放置在包括护理点在内的任何环境中，以及(3) 完成对临床样本的广泛研究，以建立性能指标。 为了在这项工作中取得成功，我们组建了一支自动化开发专家团队 诊断由具有病理学、传染病、临床专业知识的世界级顾问提供支持 微生物学、莱姆病和疏螺旋体微生物学。我们将共同努力，增强我们的能力， 提供自动化化验，最终与我们的临床合作进行盲法、多点临床研究 合伙人。在实现我们的具体目标后，我们将在ISO下开始验证和确认工作 13485质量体系，规模化生产，完成监管审批的临床试验。RAPID/LD Will 代表一项计划中的多用途面板的初步测试，该面板涵盖了一系列壁虱传播疾病(Tbd)。

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).