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

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).