Author/Editor		Stangler Herodež, Špela; Stankovič, Nastja; Zagradišnik, Boris; Erjavec Škerget, Alenka; Kokalj-Vokač, Nadja
Title		Detection of vkorc1 polymorphism
Translated title		Ugotavljanje VKORC1 polimorfizma
Type		članek
Vol. and No.		Letnik 6, št. 2
Publication year		2013
Volume		str. 47-52
ISSN		1855-5640 - Acta medico-biotechnica : AMB
Language		eng
Abstract		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.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.
Keywords		VKORC1 varfarin PCR RFLP verižna reakcija, alelno specifična polimeraza antikoagulantna terapija polimorfizem medicinska genetika