Verifikacija metode za određivanje polomorfizma gena Q192R pon 1 u bolesnika sa stenozom perifernih arterija

master's thesis

• Title (english): Verification of the method to determine the polymorphism Q 192R pon 1 gene in patients with peripheral arterial stenosis
• Author: Maja Vodopija
• Mentor: Mirjana Mariana Kardum Paro (mentor)
• Mentor: Inga Marijanović (mentor)
• Committee member: Inga Marijanović (predsjednik povjerenstva)
• Committee member: Mirjana Mariana Kardum Paro (član povjerenstva)
• Committee member: Dubravka Hranilović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2012-03-09, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Molekularno genske metode su visokospecifične metode kojima se izravno ili neizravno na razini nukleinske kiseline utvrđuje prisustvo ili odsustvo mutacija, broj kopija gena ili broj ponavljajućih jedinica. Kako je za verifikaciju ili procjenu prihvatljivosti molekularno genskih metoda neophodno sagledati cjelokupan laboratorijski proces, mnogi od ovih postupaka i radnih protokola su danas standardizirani i dostupni u obliku preporuka Nacionalnog odbora za kliničko- laboratorijske standarde (od engl. Clinical and Laboratory Standards Institute, CLSI) ili smjernica dobre laboratorijske prakse, a njihovo usvajanje je preduvjet primjene neke od molekularno genskih metoda u kliničke ili znanstveno-istraživačke svrhe. Glavni cilj istraživanja je bila verifikacija, odnosno procjena prihvatljivosti korištene molekularno genske metode u kliničke ili znanstveno-istraživačke svrhe. Specifični cilj je bio istraživanje polimorfizma Q192R u kodirajućem području gena pon1 u bolesnika sa stenozom perifernih arterija. Genetski polimorfizam paraoksonaze na kodonu 192 (Gln→Arg) je supstrat-specifičan, a navodi iz literature ukazuju na to da bi mogao biti neovisni čimbenik rizika za razvoj ateroskleroze. Metoda za određivanje polimorfizma Q192R gena pon1 (PCR-RFLP) je zadovoljila sve postavljene kriterije analitičke kvalitete prema biološkim, stručnim i drugim klinički relevantnim kriterijima. Rutinskim određivanjem polimorfizma Q192R gena pon1 u bolesnika sa stenozom perifernih arterija i kontrolnoj skupini ispitanika utvrđeno je da ne postoji statistički značajna razlika u frekvenciji genotipa i alela između kontrolne skupine i skupine bolesnika.

Abstract (english)

Molecular genetic methods are highly specific methods that can directly or indirectly on the level of nucleic acid determine the presence or absence of mutations, gene copy number or the number of repeating units in genome. It is necessary for the verification or assessment of acceptability for molecular genetic methods to examine the entire laboratory process eligibility, many of these procedures and working protocols are standardized and are available today in the form of recommendations of the National Committee for Clinical Laboratory Standards (NCLSI) and guidelines for good laboratory practice so their adoption is a prerequisite for applying some of the molecular genetic methods in the clinical or scientific research purposes. The main goal of this research was the verification, assessment of acceptability used molecular genetic method in the clinical or scientific research purposes. The specific aim was to investigate the Q192R polymorphism in the coding gene pon1 in patients with peripheral arterial stenosis. Genetic polymorphism of paraoxonase in codon 192 (Gln → Arg) is a substrate-specific, and references from the literature suggest that it could be an independent risk factor for developing atherosclerosis. Used (PCR-RFLP) method for determination of gene pon1 Q192R polymorphism has met all criteria set by biological analytical quality, professional and other clinically relevant criteria. Routine measurement of gene pon1 Q192R polymorphisms in patients with peripheral arterial stenosis and control group patients showed no statistically significant difference in genotype and allele frequencies between controls and patient groups.

• Keywords: verifikacija metode; PCR; RFLP; ateroskleroza; gen pon1; Q192R polimorfizam
• Keywords (english): verification of the method; PCR; RFLP; pon1 gene; Q192R polymorphism
• Language: croatian
• URN:NBN: urn:nbn:hr:217:201871
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-02 12:22:18