www.ksmf.org/arhimed   arhimed@ksmf.org   [5/10/2025 6:18:28 AM] http://www.ksmf.org/arhimed/poglej.asp?id=54 Identifikacija borelij, ki povzrocajo bolezen erythema migrans Avtor: Martina Munda Mentor: prof. dr. Marija Gubina, dr. med Somentor: asist. dr. Eva Ružic-Sabljic, dr. med Izhodišca: Lymska borelioza je razširjena v Evropi, Severni Ameriki in Aziji. V Sloveniji je bolezen endemska in je hkrati najpogostejša bolezen, ki jo prenašajo klopi. Kaže se z zelo razlicnimi klinicnimi slikami, prizadeti so lahko razlicni organi in organski sistemi. Najpogostejša klinicna slika je erythema migrans. Lymsko boreliozo povzrocajo razlicne vrste borelij, ki jih skupaj imenujemo Borrelia burgdorferi sensu lato (sensu lato = v širšem smislu). Namen: Namen naloge je, da iz kužnin bolnikov s klinicno sliko erythema migrans izoliramo borelije in potrdimo etiologijo kožne spremembe. Borelije želimo identificirati na nivoju vrste in s tem ugotoviti prisotnost razlicnih vrst borelij v slovenskem prostoru. Med njimi želimo ugotoviti tiste, ki povzrocajo klinicno sliko erythema migrans. Epidemiologijo želimo primerjati s tisto v evropskem prostoru in ZDA. Metode: Iz biopticnih vzorcev bolnikov s kožno spremembo erythema migrans smo izolirali in kultivirali borelije. S pomocjo metode vgraditve v gelu smo izolirali borelijsko DNA, ki smo jo nato obdelali z restrikcijskim encimom MluI. Restrikcijske odseke smo locili z elektroforezo v utripajocem elektricnem polju (PFGE). Seve smo analizirali in uvrstili v borelijske vrste glede na polimorfizem dolžine restrikcijskih fragmentov (RFLP). Rezultati: Od 30 sevov, ki smo jih izolirali, smo jih 24 (80%) uvrstili v vrsto B. afzelii in 6 (20%) v vrsto B. garinii. Noben sev ni ustrezal vrsti B. burgdorferi sensu stricto (sensu stricto = v ožjem pomenu). Sevi B. afzelii so kazali homogenost znotraj vrste, sevi B. garinii pa heterogenost, tako da smo jih razvrstili v razlicne skupine znotraj vrste. Zakljucki: Z izolacijo borelij iz biopticnih vzorcev smo potrdili borelijsko etiologijo klinicne slike erythema migrans. Kožno spremembo erythema migrans v Sloveniji povzrocata vsaj dve vrsti borelij in sicer B. afzelii in B. garinii. V naši raziskavi B. burgdorferi sensu stricto nismo uspeli izolirati. Najpogostejši povzrocitelj klinicne slike erythema migrans je B. afzelii. Pojavnost borelijskih vrst, ki smo jo zasledili v naši raziskavi, je primerljiva s pojavljanjem posameznih vrst borelij pri klinicni sliki erythema migrans v Evropi. Pri B. garinii ugotavljamo heterogenost znotraj vrste na zelo majhnem geografskem podrocju «» [Abstract / English version] Molecular characterization of borrelia, the etiologic agent of erythema migrans Author: Martina Munda Mentor: prof. dr. Marija Gubina, dr. med Co-mentor: asist. dr. Eva Ružic-Sabljic, dr. med Background: Lyme boreliosis is a widespread disease, common in Europe, North America and Asia. Lyme boreliosis is an endemic and the most common tick-borne disease in Slovenia. Signs and symptoms are various, several organs and organic systems can be affected. The most common clinical manifestation is skin rash, erythema migrans. The etiologic agents of lyme boreliosis are different genospecies of Borrelia burgdorferi sensu lato. Goal: The aim of our study was to isolate Borrelia from skin samples of patients with clinical manifestation erythema migrans, which would confirm the borrelial etiology of the skin rash. We wanted to identify the genospecies of isolated strains and to investigate the presence of different Borrelia species in Slovenia as well as identify the species, which can cause skin manifestation erythema migrans. We wanted to compare borrelial presence and prevalence in Slovenia with the one in Europe and USA. Methods: We isolated and cultivated Borrelia strains from skin samples of patients with erythema migrans. Borrelial DNA molecule was isolated with gel-insert method. We digested the DNA with MluI restriction enzyme and separated DNA fragments with Pulsed-Field Gel Electrophoresis (PFGE). We analyzed and identified borrelial species according to restriction fragments lenght polymorphism (RFLP). Results: We analyzed 30 isolates. Twenty four (80%) of them were identified as B. afzelii and 6 (20%) as B. garinii. None of the isolates were identified as B. burgdorferi sensu stricto. Isolates of B. afzelii showed homogenity within the species, but B. garinii isolates showed heterogenity and were classified in different groups within the species. Conclusions: We confirmed the borrelial etiology of clinical manifestation erythema migrans. In Slovenia, erythema migrans is caused with at least two borrelial species, B. afzelii and B. garinii. We did not succeed to isolate the species B. burgdorferi sensu stricto in our research. The most common agent of erythema migrans in our research was B. afzelii. Our results can be compared with the presence of certain borrelial species as agents of erythema migrans in Europe. We found a heterogenity within B. garinii species on a small geographic area.