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Lyme disease is a multisystem disease caused by bacteria belonging to the group Borrelia burgdorferi sensu lato. The vector that carries the infection is a tick of the genus Ixodes, that infects subsequent hosts of the spirochete during blood-meal. The varied course of Lyme disease makes it impossible to recognize it on the basis of clinical symptoms. Therefore, the diagnosis of Lyme disease is based mainly on laboratory methods, both direct (detection of the presence of DNA or infectious agent proteins in the biological material collected from the patient) and indirect (mainly serological tests). A commonly recommended approach is serodiagnosis, however, due to the time required for the body to produce specific antibodies, it is not useful in the earliest period of infection. Microbiological diagnostics also can not be used to diagnose Lyme disease in the first weeks of the disease due to its low sensitivity and long waiting time for the result. The solution seems to be molecular diagnostics based on the detection of the spirochete DNA using PCR reaction that is highly specific and sensitive. However, the effectiveness of this approach depends on many factors, therefore it is necessary to develop a standardized protocol ensuring reproducibility of results in all laboratories. 1. Introduction. 2. Genome of B. burgdorferi s.l. 3. Diagnosis of Lyme borreliosis. 4. Types of PCR reactions used in the diagnosis of Lyme disease. 5. Target genes used to DNA detection of B. burgdorferi s.l. 6. Identification of B. burgdorferi s.l. genotypes. 7. Clinical material. 8. The factors affecting the efficiency of PCR. 9. Recommendations for the use of PCR diagnostics. 10. Summary |