diplomska naloga
Nastja Stankovič (Author), Marjana Glaser (Mentor), Špela Stangler Herodež (Co-mentor)

Abstract

Cilj osebnostne medicine je izboljšati učinkovitost zdravljenja in zmanjšati pojav neželenih učinkov s prilagajanjem zdravljenja posameznemu pacientu glede na njegove genetske značilnosti. V klinični praksi se namreč bolniki na predpisani odmerek določenega zdravila pogosto ne odzovejo enako. Nekateri bolniki se na zdravljenje sploh ne odzovejo, pri nekaterih pa se pojavijo neželeni učinki. Najpogosteje uporabljeno in najbolje proučeno peroralno antikoagulacijsko zdravilo je varfarin. Za varfarin je znanih veliko stranskih učinkov (npr. krvavenje), ki so odvisni od višine odmerka. Odmerjanje varfarina je izredno individualizirano in dnevni odmerek znaša od 0,5 do 50 mg. Zdravljenje z varfarinom je odvisno tako od okoljskih kot genetskih dejavnikov. Raziskave poročajo, da je višina odmerka varfarina oz. rezistenca na varfarin povezana z mutacijo na genu VKORC1, ki nosi zapis za encim vitamin K epoksid reduktazni kompleks 1, ki je tarčni encim, na katerega deluje varfarin. Namen diplomskega dela je bil na vzorcu 441 preiskovancev splošne populacije določiti frekvenco enonukleotidnega polimorfizma oz. SNP (single nucleotid polymorphism) v genu VKORC1-1639G>A (rs9923231) s pomočjo enostavne, hitre in predvsem ekonomsko sprejemljive analizne metode. Za genotipizacijo posameznega preiskovanca smo tako izbrali metodo verižne reakcije s polimerazo/polimorfizem dolžin restrikcijskih fragmentov (PCR + RFLP) in detekcijo na agaroznem gelu. Metodo PCR + RFLP smo preverili z alelno specifično verižno reakcijo s polimerazo in opravili natančno analizo rezultatov. Ugotovili smo, da se rezultati dobljeni z metodo alelno specifične verižne reakcije s polimerazo popolnoma ujemajo s tistimi, ki smo jih dobili s pomočjo metode PCR + RFLP. Izkazalo se je, da alelno specifična verižna reakcija s polimerazo omogoča detekcijo polimorfizma gena VKORC1-1639G>A na zelo enostaven, hiter in predvsem cenovno sprejemljiv način. Dobljene ugotovitve diplomskega dela so osnova za nadaljnje raziskave prenosa te tehnologije v klinično prakso.

Keywords

gen, polimorfizem;antikoagulacijsko zdravljenje;varfarin;VKORC1 gen, mutacija;polimeraza;personalizirana medicina;

Data

Language: Slovenian
Year of publishing:
Source: Maribor
Typology: 2.11 - Undergraduate Thesis
Organization: UM - University of Maribor
Publisher: [N. Stankovič]
UDC: 577.112.38+575.17(043.2)
COBISS: 4049215 Link will open in a new window
Views: 3795
Downloads: 266
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Other data

Secondary language: English
Secondary title: Detection of VKORC1 polymorphism: comparison of polymerase chain reaction/restriction fragment lenght polymorphism (PCR + RFLP) with allele specific polymerase chain reaction
Secondary abstract: The goal of personalized medicine is to improve health efficacy and minimize adverse drug reactions by adapting the treatment to each patient regarding his genetic information. In clinical practice the patients do not react the same way on prescribed dose of certain medicine. Some patients are not responding at all, while others experience side effects. The most widely prescribed and best studied oral anticoagulant is warfarin. Warfarin has many side effects (e.g. bleeding) depending on the dosage amount. Warfarin dosing is very individualized and the daily requirement varies from 0,5 to 50 mg. Warfarin therapy is influenced by environmental and genetic factors. The study researches are indicating that the dosage amount of warfarin and resistance to warfarin are associated with mutation in VKORC1 gene, which encodes for enzyme vitamina K epoxide reductase complex 1. This is the target enzyme, which is affected by warfarin. The aim of my diploma thesis which is based on a sample of 441 individuals of the general population, was to determine the frequency of single nucleotide polymorphism (SNP) in the gene VKORC1-1639G>A, using a simple, rapid and economically reasonable analytic method. For genotyping each individual we chose the method of polymerase chain reaction/restriction fragment length polymorphism (PCR + RFLP) and detection on agarose gel. PCR + RFLP method was tested with allele specific polymerase chain reaction and the exact analyse of the results was made. The results we obtained with allele specific polymerase chain reaction are entirely consistent with those we obtained using PCR + RFLP method. It turned out that allele specific polymerase chain reaction allows detection of polymorphism of gene VKORC1-1639G>A in a simple, rapid and economically acceptable way. The obtained findings of my diploma thesis are the basis for further researches of transferring this technology into clinical practice.
Secondary keywords: VKORC1;warfarin;PCR;RFLP;allele specific polymerase chain reaction;
URN: URN:SI:UM:
Type (COBISS): Undergraduate thesis
Thesis comment: Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages: 51 f.
Keywords (UDC): mathematics;natural sciences;naravoslovne vede;matematika;biological sciences in general;biologija;material bases of life;biochemistry;molecular biology;biophysics;biokemija;molekularna biologija;biofizika;mathematics;natural sciences;naravoslovne vede;matematika;biological sciences in general;biologija;general genetics;general cytogenetics;splošna genetika;splošna citogenetika;
ID: 1015886