Researchers at Dartmouth Hitchcock Medical Center developed three droplet digital PCR (ddPCR) assays that may improve early and accurate detection of Lyme disease. Presented at AMP 2025, the assays target all Borrelia species, Lyme-causing Borrelia, and B. burgdorferi specifically. Validated on reference strains and clinical samples, they detected as few as 5–10 bacterial cells with high specificity. The method successfully identified active B. burgdorferi DNA in a patient whose antibody tests suggested only past exposure. By measuring bacterial DNA directly, the assays could reduce diagnostic delays and work across multiple ddPCR platforms, supporting broad laboratory adoption.