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The investigations aimed to establish the reliability of the chosen serological tests designed for the diagnosis of
Lyme borreliosis in horses. The investigations were carried out in five Horse Breeding Centres (OHK). Statistical analysis methods were used to determine sample size for particular centres: Krasne (Kr)–49, ?ack (?)–21, Walewice (W)–111, Bogus?awice (B)–17, Kozienice (K)–61. The experimental material comprised the chosen horses from which blood samples were collected in order to obtain sera. The test used for indirect immunofluorescence assay (IFA No 75941, Bio-Mérieux) is commercially designed for the investigation of human sera and thus needed a prior species adaptation and standardization; ELISA (MRL DIAGNOSTICS, No EL0400G) which was also species adapted and stardandized and ELISA commercially assigned for the examination of dog or horse sera (Die System Diagnostica GmbH Borrelia burgdorferi Veterinary ELISA No. 122.00 Genzyme Virotech GmbH). In the IFA test the highest share of positive results was obtained in respect of the sera from OHK in (K)–60.7% and then in (B)–52.9%, (?)–42.9%, (W)–40.5%, (Kr)–38.7%. In the standardized ELISA the highest percent of positive results, amounting to 33.3%, was obtained in respect of the sera from (?), and then from (W)–20.7%, (K)–11.5%, (Kr)–10.2% and (B)–5.9%. The percent of positive results obtained in the commercial ELISA also agreement on a high level: the sera originating from (W) were positive in 18.9%, from (K)–9.8%, (?)–9.5%. (B)–5.9% and (Kr)–4.1%. Both ELISAs showed high agreement although the standardized test was characterized by a greater tendency for suggesting the presence of B. burgdorferi infection and the agreement of these two ELISAs with the IFA was not so strong. The IFA showed the highest tendency for suggesting the presence of the B. burgdorferi infection, being characterized by the highest percent of false positive results.