摘要： Background: Current serological tests cannot discriminate between bactericidal B. burgdorferi antibodies from others that are merely a response to Borrelia antigenic stimulation. Objective: To develop a sensitive and convenient luminescence-based serum bactericidal assay (L-SBA) to identify serum borreliacidal activity. Study design: Prospective validation study and method comparison. Methods: Serum samples were obtained either from archives of Animal Health Diagnostic Center at Cornell University (N = 7) or a vaccination trial (N = 238). Endogenous complement-inactivated serum sample was incubated with exogenic complement and Borrelia burgdorferi ML23 pBBE22luc, which is able to process luciferin with luciferase and produce luminescence in viable Borrelia. After incubation, a light signal can be detected by using a luminometer and calculate the borreliacidal antibody titer. Results: Components of reaction mixture including spirochetes and complement from various sources and concentrations were tested to identify a reliable recipe for our complement mediated L-SBA. We also applied this L-SBA on measuring bactericidal antibody activities and calculated IC50 of serum samples from clinical collections. Furthermore, we analysed the L-SBA titers and anti-OspA antibody levels from vaccinated horses using the multiplex assays and found that there is a relationship between results generated using these two different assays. The increases of L-SBA titers correlated with increases of anti-OspA antibody titer in sera (r = 0.423). Main limitations: Immunoreactivity of commercial complement may differ from different batches. Clinical protection of borreliacidal antibody levels has not been determined. Conclusions: The L-SBA provided a sensitive and easy-operating platform for evaluation of bactericidal antibody to Borrelia burgdorferi, and we anticipated L-SBA would function well as an evaluation tool of vaccine efficiency in the future.