Novel Point-of-Care Multiplex Lyme Disease Test

DESCRIPTION (provided by applicant): Lyme disease (LD) is caused by spirochetal bacteria from the genus Borrelia. Borrelia burgdorferi is the predominant cause of Lyme disease in the United States, whereas Borrelia afzelii and Borrelia garinii are implicated in most European cases. CDC reported 27,444 cases of Lyme disease in the United State in 2007 alone. It is estimated 2.5 million LD serological tests have been done annually. In response to the solicitation from NIAID in developing novel approaches for the diagnosis of Lyme disease, and to address the limitation of current LD assays, we propose to develop a highly sensitive Point-of-Care Multiplex Hand-Held Diagnostic Device capable of detecting the presence IgM and IgG antibodies against Borrelia species proteins in human serum, human saliva and human urine. The technology of our device is based on immunoprecipitation principles similar to those found in popular pregnancy tests. Lateral-flow chromatographic and flow-through tests offer fast detection of critical components for use in POC testing. The key feature of a proposed flow- through tests to achieve high sensitivity is the ability to covalently attach antibodies or antigens to highly intensely colored, nanometer-sized colloidal gold. Our lateral flow test assay development experience has demonstrated that the assay is as sensitive as ELISA, while reducing assay time from 8hrs to 5 minutes. Another key feature of the proposed test is the collection of 14 Borrelia specific in vivo-induced, non-denatured antigens that have been shown to elicit strong antibody response in humans. These antigens will be used to capture human Borrelia antibodies presented in human serum samples. In Phase I, we will use recombinant antigens of B. burgdorferi species, in Phase II, for each antigen we will use recombinant proteins from all three Borrelia species - B. burgdoferi, B. afzelii and B. garinii to further increase sensitivity. The POC assay will be inexpensive, less than 20 per test, robust, requires only a single step, field-capable, user-friendly, and temperature stable, with no requirement for sample processing. The results can be read in 10 to 30 minutes. PUBLIC HEALTH RELEVANCE: Lyme disease (LD) is the most widespread tick-borne disease affecting more than 27,000 people in the US in 2007 alone. The current LD diagnosis tests are both time consuming and expensive. We propose to develop a high accuracy Point-of-Care test similar to common pregnancy test. It is inexpensive, less than 20 per test, robust, requires only a single step, user-friendly, and temperature stable, with no requirement for sample processing. The results can be read in 10 to 30 minutes.