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master's thesis
Predanalitički čimbenici u analizi cfDNA
Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry, 2018. urn:nbn:hr:163:519750
Hlapčić, Iva
University of Zagreb
Faculty of Pharmacy and Biochemistry
Department of biochemistry and molecular biology

Institutional repository: Repository of Faculty of Pharmacy and Biochemistry University of Zagreb

Cite this document

Hlapčić, I. (2018). Predanalitički čimbenici u analizi cfDNA (Master's thesis). Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry. Retrieved from https://urn.nsk.hr/urn:nbn:hr:163:519750

Hlapčić, Iva. "Predanalitički čimbenici u analizi cfDNA." Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, 2018. https://urn.nsk.hr/urn:nbn:hr:163:519750

Hlapčić, Iva. "Predanalitički čimbenici u analizi cfDNA." Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, 2018. https://urn.nsk.hr/urn:nbn:hr:163:519750

Hlapčić, I. (2018). 'Predanalitički čimbenici u analizi cfDNA', Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, accessed 09 November 2024, https://urn.nsk.hr/urn:nbn:hr:163:519750

Hlapčić I. Predanalitički čimbenici u analizi cfDNA [Master's thesis]. Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry; 2018 [cited 2024 November 09] Available at: https://urn.nsk.hr/urn:nbn:hr:163:519750

I. Hlapčić, "Predanalitički čimbenici u analizi cfDNA", Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, 2018. Available at: https://urn.nsk.hr/urn:nbn:hr:163:519750

Metadata
TitlePredanalitički čimbenici u analizi cfDNA
Title (english)Preanalytical factors in cfDNA analysis
AuthorIva Hlapčić
MentorSandra Šupraha Goreta (mentor)
Committee memberSandra Šupraha Goreta (predsjednik povjerenstva)
Committee memberOlga Gornik (član povjerenstva)
Committee memberNada Vrkić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Pharmacy and Biochemistry
(Department of biochemistry and molecular biology)
Zagreb
Defense date and country2018-02-26, Croatia
Scientific / art field,
discipline and subdiscipline		BIOMEDICINE AND HEALTHCARE
Pharmacy
Medical Biochemistry
Abstract
Karcinom dojke najučestalije je maligno oboljenje u Hrvatskoj, a dijagnoza se vrlo često postavlja kasno što utječe na daljnji tijek bolesti. Važno je poboljšati laboratorijsku dijagnostiku da bi se otkrile i standardizirale metode koje bi omogućile specifično i rano postavljanje dijagnoze. Biomarkeri, koji se danas koriste, naišli su na svoja ograničenja, stoga se želi unaprijediti laboratorijski proces u ranom otkrivanju karcinoma dojke kao i svih ostalih malignih bolesti. Slobodna cirkulirajuća DNA, cfDNA, potencijalni je biomarker karcinoma budući da se velikim dijelom u cirkulaciju otpušta iz tumorskog tkiva. Prednosti, u odnosu na trenutno poznate biomarkere, su u tome što je metoda neivazivna jer je potreban samo uzorak krvi, daje uvid u trenutni molekularni status što omogućuje personalizirani pristup, bolje praćenje statusa bolesti i terapije. Ovim radom željelo se ispitati kako predanalitička faza utječe na kvalitetu i kvantitetu cf DNA. Koristili su se uzorci plazme i seruma pacijentica s karcinomom dojke sa i bez metastaza te kontrolni zdravi uzorci. Uzastopnim odmrzavanjem i zamrzavanjem uzoraka vidjelo se da se time narušava integritet DNA pa tako i kvaliteta te kvantiteta izoliranog materijala. Primjenom dviju vrsta kolona za izolaciju, ustanovljeno je da je za izolaciju cf DNA potrebno koristiti specifične kolone namijenjene za cfDNA da bi doprinos bio veći jer genomska DNA nepovoljno utječe na izolaciju cfDNA. Nadalje, različiti stadiji bolesti utječu na integritet cfDNA što otežava izolaciju. Dodatno se javlja problem povećane koncentracije cfDNA kod zdravih pojedinaca što znači da postoje fiziološka stanja popraćena povećanom koncentracijom cfDNA. Svi navedeni rezultati ukazuju na potrebu za specifičnim genetičkim analizama.
Abstract (english)
Breast cancer is the most common malignant disease in Croatia and it is often diagnosed too late which affects the course of disease. It is important to improve laboratory diagnostics to detect and standardize methods that would allow a specific and early diagnosis. The biomarkers, that are used nowadays, have their limitations, so there is an urge in advancing the laboratory process of early detection of breast cancer as well as other malignant diseases. Free circulating DNA, cfDNA, is a potential cancer biomarker since it is released into the circulation from tumor tissue. The advantages over the currently-known biomarkers are that the method is non invasive because only blood sample is needed, it provides an insight into the molecular status at theparticular moment and enables personalized medicine, better monitong of disease progression and therapy. This research investigated how the pre-analytical phase affects the quality and quantity of cfDNA. Samples of plasma and serum from breast cancer patients with and without metastases were used as the samples from healthy controls. Several freezing and defrosting affected in a way that the quality and quantity of DNA were poor. Two different colons for isolation were used. It was found that isolation of cfDNA requires specific columns intented for specific isolation of cfDNA so that DNA yield would be bigger and more stable despite the presence of genomic DNA. Moreover, different stages of the disease affect DNA integrity which makes isolation more difficult. In addition, there is a problem of increased cfDNA concentration in healthy individuals which means there are some physiological states with elevated cfDNA concentration. All these results suggest specific genetic analysis.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:163:519750
Study programmeTitle: Medical biochemistry
Study programme type: university
Study level: integrated undergraduate and graduate
Academic / professional title: magistar/magistra medicinske biokemije (magistar/magistra medicinske biokemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2018-12-13 16:03:26