The aim of the study was the assessment of changes in the serum phospholipid profile patients with Lyme neuroborreliosis (NB), asymptomatic people after frequent tick bites and patients after long-term multidrug therapy against B.burgdorferi. LC-QTOF-MS/MS platform was used to identify changes in serum phospholipid profile of 37 persons. The results demonstrate differences PL profile among patients with Lyme borreliosis (LB), people frequently exposed to tick bites and patients treated with long-term multidrug therapy compared to healthy subjects. Significant differences in SM, LPC, PI, and PC content of NB patients discriminate this group of patients from the other groups, have potential diagnostic value, and can be used for the development of more effective diagnostic tools. The finding that the phospholipid profile of foresters is similar to healthy people, suggests the existence of adaptive mechanisms that are currently difficult to explain but are interesting from the perspective of future research.