Project Summary/Abstract Lyme disease (LD) is the most common vector-borne illness in the United States and represents a considerable diagnostic challenge. 1-3 The current benchmark for LD diagnosis is the standard two-tiered test (STTT), although the modified two-tier test (MTTT) is rapidly becoming an acceptable alternative. 6 These assays are highly specific but have poor sensitivity, especially early in the infection. 6, 7 Traditional testing is limited by each patient's heterogenous immune response to Borrelia burgdorferi (Bb) antigens. While the paired ELISAs of the MTTT are sensitive and simple to perform, some patients respond weakly to screening reagents. The STTT addresses variation in humoral response by combining an initial ELISA with a confirmatory IgG/IgM Western Blot (WB), yet WB sensitivity is modest against any single antigen. This is especially true with IgM results, which also lack the specificity enhancement associated with class switching. 8 Given these limitations, the ideal serologic assay will combine features of ELISA and WB, but will provide quantitative and specific results against all major LD antigens. Using current technologies, this approach is cost prohibitive. Ciencia Inc. has developed a low-cost, highly-sensitive, fluorescence-based assay platform that supports exactly this type of quantitative multi-antigen serologic Lyme assay. 4, 9, 10 Dr. Cady at SUNY Poly has used our grating-coupled fluorescent plasmonics (GC-FP) assay platform to define antibody responses against a suite of Bb antigens. This preliminary work demonstrates superior sensitivity/specificity compared to STTT (90%/100% vs 60%/100%; n=34 validated samples) and an earlier time to result. Our approach offers a linear response across the full dynamic range, a rapid time to result, low reagent costs. Antigen spots may be added for a negligible cost, so a broader tickborne disease (TBD) panels could be performed for the price of one ELISA. With this Phase I STTR, Ciencia will partner with Dr. Cady (SUNY Polytechnic Institute) and Dr. Strle (Wadsworth Center) to build upon our preliminary self-funded exploration of Lyme serodiagnostics. We aim to validate our preliminary results using a larger set of well-characterized LD samples and appropriate controls from the CDC and NY State DoH, including an estimate of time to positivity using samples collected longitudinally. In parallel, Ciencia aims to improve sample handling and overall ease of use by implementing a cartridge-based system and automated processing that will make GC-FP a viable alternative to methods such as the STTT and MTTT, both of which are cumbersome and time consuming. The ultimate goal is to provide a rapid, easy-to-use, and highly accurate test that can be used outside traditional clinical laboratories. Based on the success of Phase I, the Phase II effort would optimize the test for diagnosis of Lyme disease caused by European Borrelia strains and expand it to other tickborne diseases. Phase II would also permit design refinements and initiation of the FDA approval process.

Project Narrative Lyme Disease diagnosis is hindered by low titers of clinically relevant antibodies early in infection and complex multi-tier testing protocols. With this project, Ciencia and collaborators respond to the “Notice of Special Interest: Advancing Research for Tickborne Diseases” by further validating our Lyme serodiagnostics assay and by developing an easy-to-use system for sample handling and data interpretation. Through partnership with SUNY Polytechnic Institute and the Wadsworth Center (NY State Dept. of Health), Ciencia aims to develop a next generation assay platform that can bring rapid, low-cost, easy-to-use, and high-quality diagnostics outside traditional testing sites.