20.500.12556/DKUM-88256 Detection of vkorc1 polymorphism comparison of polymerase chain reaction/restriction fragment length polymorphism (pcr + rflp) with allele-specific polymerase chain reaction Ugotavljanje VKORC1 polimorfizma primerjava metod verižne reakcije s polimerazo/polimorfizma dolžin restrikcijskih fragmentov (PCR + RFLP) z alelno specifično verižno reakcijo s polimerazo Purpose: The VKORC1 polymorphism is an important genetic factor affecting warfarin dose requirement. Patients require different warfarin doses in order to achieve the target therapeutic anticoagulation. The aim of our study was to determine the frequency of single nucleotide polymorphisms (SNP) in the VKORC1gene in the general population, using a simple, rapid, and economical method. Methods: For genotyping, the restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR) amplified DNA was used and compared to allele specific polymerase chain reaction. We genotyped 441 DNA samples obtained from the healthy general population in North Eastern Slovenia. Genotypes for the tested group were evaluated to determine whether the population followed the Hardy Weinberg equilibrium. The genotypes and allele frequencies were calculated. Results: The results obtained using the allele specific polymerase chain reaction were consistent with those obtained using the PCR + RFLP method. The G allele frequency (0.62) was higher than the A allele frequency (0.38) in the general population from North Eastern Slovenia. Conclusions: The PCR+RFLP method involved additional manipulation of the PCR products at the expense of analysis time, consumption of reagents and equipment. The allele specific polymerase chain reaction was a simple and rapid method for the detection of SNP in theVKORC1 gene, and is available in any laboratory with the minimum of equipment and reagents required. Namen: VKORC1 polimorfizem je pomemben genetski dejavnik, ki vpliva na zahtevan odmerek varfarina pri bolnikih, ki potrebujejo različne odmerke varfarina za doseganje antikoagulacijskega terapevtskega učinka. Cilj naše raziskave je bil na vzorcu splošne populacije ugotoviti frekvenco enonukleotidnega polimorfizma (SNP) v VKORC1 genu na preprost, hiter in ekonomičen način. Metode: Za genotipizacijo smo uporabili metodo verižne reakcije s polimerazo oz. polimorfizem dolžin restrikcijskih fragmentov (PCR/RFLP), ki smo jo primerjali z metodo alelno specifične verižne reakcije s polimerazo. Genotipizirali smo 441 vzorcev genomske DNA preiskovancev iz severovzhodne Slovenije. Ocenili smo, ali je izbrana skupina v Hardy Weinbergovem ravnovesju, ter izračunali posamezne genotipe in frekvenco alelov. Rezultati: Rezultati, pridobljeni z alelno specifično verižno reakcijo s polimerazo, so se popolnoma ujemali z rezultati, pridobljenimi z metodo PCR/RFLP. Frekvenca alela G (0,62) je višja kot frekvenca alela A (0,38) v testirani skupini iz severovzhodne Slovenije. Zaključek: Izkazalo se je, da PCR/RFLP metoda vključuje dodatne korake na račun časa analize, porabe reagentov in opreme. Na podlagi rezultatov zaključujemo, da alelno specifična verižna reakcija s polimerazo omogoča odkrivanje SNP v VKORC1 genu na bolj enostaven, hitrejši in cenovno bolj sprejemljiv način. VKORC1 varfarin PCR RFLP alelno specifična verižna reakcija s polimerazo VKORC1 warfarin PCR RFLP allele–specific polymerase chain reaction true true false Angleški jezik Slovenski jezik Znanstveno delo 2024-04-12 10:19:13 2024-04-12 10:19:14 2026-03-19 12:57:11 0000-00-00 00:00:00 2013 0 0 str. 47-52 št. 2 Letn. 6 2013 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2013-01-01 575 4863039 1855-5640 http://www.actamedbio.mf.uni-mb.si/05_11id_amb_69_12.pdf 1 5a850bb4-f8a5-11ee-a0d6-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=178392 05.pdf 05.pdf 1 1A0FD9965330E6A10F531EF68D18EA59 c84969d28fe361fbfcefb7b47a3ab6fe3540dcc578a83d96e9492e69e4011c95 9c07a02d-f8a5-11ee-a0d6-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=178394 Medicinska fakulteta Acta medico-biotechnica 0 0 0