Provided herein is a method of generating a
binding pattern from a sample obtained from a subject. In some embodiments, the method includes detecting binding of antibodies from the sample to an array of peptides that each comprise at least subsequences of B. burgdorferi
protein sequences listed in SEQ ID NOS: 1-45 to produce a detected binding
data set, and identifying one or more binding patterns in the detected binding
data set. In some embodiments, the method includes classifying the
binding pattern as indicative of the subject having
Lyme disease, as indicative of the subject not having
Lyme disease, or as indicative of the subject having a non-Lyme febrile
disease. Provided also herein are other methods and related reaction mixtures, compositions, kits, devices, systems, and computer readable media.