The aim was to determine the prevalence of serum antibodies to tick-borne pathogens and other infectious agents in patients with lymphadenopathy. Materials and methods. 68 patients with lymphadenopathy aged 20 to 63 years who received outpatient or inpatient treatment were followed up at Ternopil University Hospital and Ternopil Regional Clinical Dermatological and Venereological Dispensary. There were 41 women (60.3 %) and 27 men (39.7 %). Two-stage serological detection (ELISA and immunoblot) was used for diagnosis of Lyme borreliosis (LB). Indirect immunofluorescence, including the BIOCHIP multiplex indirect immunofluorescence technique, was used for serological identification of human granulocytic anaplasmosis (HGA) and other infectious diseases, which could be associated with lymphadenopathy. Results. In total, 45.6 % of 68 enrolled patients were positive for the presence of specific antibodies IgM and/or IgG against B. burgdorferi s. l. Further examination demonstrated the presence of specific serum anti-A. phagocytophilum antibodies IgM and/or IgG in 33 patients with lymphadenopathy. It was found that 24.2 % of involved patients were positive for the presence of at least one type of anti-A. phagocytophilum antibodies; 12.1 % had positive or intermediate results only for B. burgdorferi s. l.; 60.6 % of patients were positive for the presence of antibodies against both pathogens. 46.9 % of patients were seropositive for EBV-CA, 40.6 % – for Parainfluenza virus type 1, 37.5 % – for HHV-6, 34.4 % – for HSV-1, 31.3 % – for EBV-EA and Toxoplasma gondii, 6.3 % – for Coxsackievirus type A9 and HIV-2. Conclusions. One-third of all involved patients with lymphadenopathy were positive for antibodies to B. burgdorferi s. l., causa tive agent of Lyme borreliosis. Co-infection of Lyme borreliosis and human granulocytic anaplasmosis was established in more than half of the patients. Besides, specific antibodies to 18 causative agents of other infectious diseases were detected with varying rate. Almost half of positive results detected were the antibodies to parainfluenza virus type 1 and Epstein–Barr virus.