Until 10 years ago, Lyme disease (LD) was mostly unknown in USA, although some of its clinical manifestations have been reported in Europe since 1909. The first LD cases (actually more than 50 in the same area) in USA were discovered in 1975; and its etiology (Borrelia burgdorferi) and mode of infection (tick bites) were determined a few years later. With this information, defined clinical features and serologic tests available, LD has been reported in increasing number worldwide. LD has 3 major clinical stages lasting 2 years or longer, with a number of its symptoms and signs very similar to many other diseases. It can be cured with antibiotics. Serology plays a very important role in diagnosis and management of LD. The serologic tests available are designed for clinical laboratories, which are often not easily available in suburban and rural areas were LD occurrs. A simple, quick, safe and in-expensive serologic test for LD would allow LD diagnosed in physician office or small clinical laboratories and specifically treated earlier and more efficiently. The objective of this research project is to develop such a test. The test will use a simple plastic device and require three sequential additions of two drops of patient blood sample, enzyme-antibody conjugate and substrate, with a few drops of wash solution in between. The test will take less than 15 minutes and the results will be read visually. Phase I will concentrate on: 1) growing B. burgdorferi organisms in culture; 2) accumulation of clinical serum samples from LD and other diseases; 3) preparation of various antigens; 4) testing these antigens in crude and eventually final plastic device called flex well; 5) design and making an innovative plastic device which will be used to coat antigens and receive test reagents in the test; 6) study of all assay parameters which will eventually lead to the best or optimum test procedure; and 7) finalization of test procedure with testing available clinical serum samples. Phase II will do field evaluation, refinement of test, and scale-up production design.