Vrednovanje sustava LightCycler PRO System za genotipizaciju polimorfizama FV Leiden, MTHFR i PAI-1

master's thesis

• Title (english): Evaluation of the LightCycler PRO System for genotyping FV Leiden, MTHFR and PAI-1 polymorphisms
• Author: Filip Markić
• Mentor: Mario Štefanović (mentor)
• Committee member: Mario Štefanović (predsjednik povjerenstva)
• Committee member: Anita Somborac Bačura (član povjerenstva)
• Committee member: Ivana Ćelap (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; Zagreb
• Defense date and country: 2024-10-07, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Trombofilija je skup poremećaja koji povećavaju sklonost organizma stvaranju krvnih ugrušaka (tromba), što
može dovesti do ozbiljnih zdravstvenih komplikacija kao što su duboka venska tromboza i plućna embolija.
Nasljedni rizični čimbenici uključuju genske polimorfizme faktor V Leiden (FV G1691A), PAI-1 5G/4G i
MTHFR C677T. U Kliničkom zavodu za kemiju Kliničkog bolničkog centra Sestre milosrdnice instaliran je
novi analitički sustav LightCycler® PRO (Roche Diagnostics GmbH, Basel, Švicarska) koji bi se koristio za
genotipizaciju navedenih polimorfizama. Cilj ovog rada bio je vrednovati analitički sustav LightCycler® PRO
validacijom analitičkih karakteristika mjernih postupaka genotipizacije FV G1619A, PAI-1 5G/4G i MTHFR
C677T metodom lančane reakcije polimeraze uz analizu krivulje taljenja prije korištenja u rutinskom radu.
Ispitani su analitička nepreciznost, analitička osjetljivost, analitička specifičnost, analitička netočnost,
usporedivost drugom metodom i granica detekcije. Svi ispitani parametri u potpunosti su zadovoljili unaprijed
definirane analitičke kriterije kvalitete i stoga se ispitani mjerni postupci mogu prihvatiti za rutinski rad.

Abstract (english)

Thrombophilia is a collection of disorders which increase an organism's tendency to form blood clots, which can
lead to serious health complications like deep venous thrombosis and pulmonary embolism. Hereditary risk
factors include gene polymorphisms for factor V Leiden (FV G1691A), PAI 5G/4G, and MTHFR C677T. New
analytical system for genotyping aforementioned polymorphisms, LightCycler® PRO (Roche Diagnostics
GmbH, Basel, Švicarska), was installed in the Clinical Department of Chemistry University Hospital Center
Sestre milosrdnice. This study aimed to evaluate new analytical system LightCycler® PRO by validating
analytical characteristics of genotyping polymorphisms FV G1691A, PAI-1 5G/4G, and MTHFR C677T using
real-time polymerase chain reaction with melting curve analysis. Analytical imprecision, analytical sensitivity,
analytical specificity, analytical inaccuracy, repeatability using another method, and limit of detection were
examined. All parameters fully met the pre-defined analytical quality criteria and therefore the evaluated methods
can be accepted for routine work.

• Keywords: factor V Leiden; MTHFR; PAI-1; LightCycler® PRO; genotipizacija polimorfizama
• Keywords (english): Factor V Leiden; MTHFR; PAI-1; LightCycler® PRO; polymorphism genotyping
• Language: croatian
• URN:NBN: urn:nbn:hr:163:835017
• Study programme: Title: Medical biochemistry - integrated undergraduate and graduate - university study; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: sveučilišni magistar medicinske biokemije i laboratorijske medicine (sveučilišni magistar medicinske biokemije i laboratorijske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2025-01-29 13:41:55