20.500.12556/RUL-171795 Določanje prisotnosti patogenih ekspanzij GAA v genu FGF14 pri bolnikih z ataksijo Characterization of the FGF14 gene expansion in patients with ataxia Spinocerebelarna ataksija 27 (SCA27), je bila pred nedavnim vzročno povezana z dolgimi ponovitvami nukleotidnega motiva GAA v prvem intronu gena FGF14 (fibroblastni rastni faktor 14). Namen naše raziskave je bil določiti prisotnost patogenih ponovitev GAA v genu FGF14 pri slovenskih bolnikih z nepojasnjeno ataksijo. V raziskavo smo vključili 103 takšnih bolnikov. Za genetsko dokazovanje smo uporabili verižno reakcijo s polimerazo (PCR), ter prisotnosti ponovitev GAA dodatno potrdili s pomočjo Sangerjeve sekvenčne analize. V prvi fazi smo s PCR določili dolžino ponovitev, kar je bilo uspešno pri 101 (98 %) bolnikov. Nato smo pri bolnikih, ki so na vsaj enem alelu pokazali prisotnost fragmentov večjih od normalnega območja ponovitev GAA, izvedli še sekvenčno analizo za potrjevanje ponovitev GAA. Pri petih (4,9 %) bolnikih smo identificirali heterozigotne patogene ponovitve z dolžino nad 300 ponovitev, kar potrjuje prisotnost GAA-FGF14 ataksije, pri dveh (2,0 %) bolnikih pa smo dokazali alele s ponovitvami v območju zmanjšane penetrance (med 250 in 300 ponovitev). Skupno so bile ponovitve GAA verjeten vzrok ataksije pri sedmih (6,9 %) slovenskih bolnikov z nepojasnjeno ataksijo, kar predstavlja podoben delež kot v drugih populacijah z znano prevalenco te bolezni. Naša študija potrjuje, da so patogene ponovitve GAA v genu FGF14 prisotne tudi v slovenski populaciji bolnikov z ataksijo. Uporabljeni diagnostični metodi sta se izkazali zanesljivi in učinkoviti pri odkrivanju teh sprememb, kar pomembno prispeva k izboljšanju genetske diagnostike ter razumevanja molekularnega ozadja ataksij. Spinocerebellar ataxia 27 (SCA27) has recently been associated with long GAA expansions in the first intron of the FGF14 (fibroblast growth factor 14) gene. The aim of our study was to determine the presence of pathogenic GAA expansions in the FGF14 gene in Slovenian patients with unexplained ataxia. In our study, we included 103 patients with genetically unexplained ataxia. For genetic testing, we used polymerase chain reaction (PCR) and confirmed the presence of GAA repeats with Sanger sequencing analysis. In the first phase, we used PCR to determine the repeat length, which was successful in 101 (98 %) patients. With patients who showed fragments larger than the normal GAA repeat range on at least one allele, we performed additional sequencing analysis. We identified heterozygous pathogenic expansions longer than 300 repeats in five patients (4,9 %), confirming the presence of GAA-FGF14 ataxia in these patients. In two patients (2,0 %), we detected alleles with repeats in the reduced penetrance range (between 250 and 300 GAA repeats). In total, GAA repeat expansions were the likely cause of ataxia in seven (6,9 %) Slovenian patients with unexplained ataxia, which is comparable to the prevalence reported in other populations. Our study confirms that pathogenic GAA expansions in the FGF14 gene are also present in the Slovenian patients with ataxia. The diagnostic methods used proved to be reliable and effective in detecting these mutations, significantly contributing to the improvement of genetic diagnostics and the understanding of the molecular basis of ataxia. Spinocerebelarna ataksija FGF14 PCR Sanger sekvenciranje ekspanzije Spinocerebellar ataxia FGF14 PCR Sanger sequencing expansions true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-09-03 07:15:36 2025-09-03 07:15:44 2025-09-20 03:52:23 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 262033 247564035 50667.pdf 50667.pdf 1 E9F5F886992FB922C68F2DFD5AD8239C 1d297184c32ae8b8d0ae4cf705f910b36b0e2020d44ab3726afbe6be12db8267 e5592eb9-8884-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=215285 Biotehniška fakulteta 0 0 0