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master's thesis
Usporedba kvantifikacije DNA virusa Epstein-Barr primjenom dvaju testova lančane reakcije polimerazom u stvarnom vremenu
Zagreb: University of Zagreb, Faculty of Science, 2024. urn:nbn:hr:217:180479
Listeš, Ivana
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Listeš, I. (2024). Usporedba kvantifikacije DNA virusa Epstein-Barr primjenom dvaju testova lančane reakcije polimerazom u stvarnom vremenu (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:180479

Listeš, Ivana. "Usporedba kvantifikacije DNA virusa Epstein-Barr primjenom dvaju testova lančane reakcije polimerazom u stvarnom vremenu." Master's thesis, University of Zagreb, Faculty of Science, 2024. https://urn.nsk.hr/urn:nbn:hr:217:180479

Listeš, Ivana. "Usporedba kvantifikacije DNA virusa Epstein-Barr primjenom dvaju testova lančane reakcije polimerazom u stvarnom vremenu." Master's thesis, University of Zagreb, Faculty of Science, 2024. https://urn.nsk.hr/urn:nbn:hr:217:180479

Listeš, I. (2024). 'Usporedba kvantifikacije DNA virusa Epstein-Barr primjenom dvaju testova lančane reakcije polimerazom u stvarnom vremenu', Master's thesis, University of Zagreb, Faculty of Science, accessed 03 May 2024, https://urn.nsk.hr/urn:nbn:hr:217:180479

Listeš I. Usporedba kvantifikacije DNA virusa Epstein-Barr primjenom dvaju testova lančane reakcije polimerazom u stvarnom vremenu [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2024 [cited 2024 May 03] Available at: https://urn.nsk.hr/urn:nbn:hr:217:180479

I. Listeš, "Usporedba kvantifikacije DNA virusa Epstein-Barr primjenom dvaju testova lančane reakcije polimerazom u stvarnom vremenu", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2024. Available at: https://urn.nsk.hr/urn:nbn:hr:217:180479

Metadata
TitleUsporedba kvantifikacije DNA virusa Epstein-Barr primjenom dvaju testova lančane reakcije polimerazom u stvarnom vremenu
Title (english)Comparison of Epstein-Barr virus DNA quantification using two real-time polymerase chain reaction assays
AuthorIvana Listeš
MentorSnježana Židovec Lepej (mentor)
Committee memberSnježana Židovec Lepej (predsjednik povjerenstva)
Committee memberPetra Korać (član povjerenstva)
Committee memberMaja Matulić (član povjerenstva)
Committee memberSandra Hudina (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2024-02-28, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Virus Epstein-Barr (EBV) onkogeni je virus koji pripada porodici Orthoherpesviridae i rodu Lymphocryptovirus čiji se genom sastoji od dvolančane molekule DNA koja sadrži oko 172 kb. Virus EBV umnožava se u epitelnim stanicama, B-limfocitima, T-limfocitima i NK-stanicama, a opisana su dva glavna tipa replikacijskog ciklusa EBV-a; latentni i litički ciklus. Cilj ovog istraživanja bio je usporediti broj kopija DNA EBV-a u uzorcima periferne krvi određenih primjenom dvaju standardiziranih kvantitativnih PCR testova u stvarnom vremenu te utvrditi jesu li rezultati procjene kinetike virusne replikacije usporedivi i primjenjivi u biološkim i biomedicinskim istraživanjima. Rezultati detekcije DNA EBV-a u 100 uzoraka periferne krvi qPCR testovima u stvarnom vremenu LightMix LightCycler 2.0 i LightMix LightCycler 480 II s adaptiranim protokolom bili su podudarni u 99 od 100 uzoraka (Kohenov kappa koeficijent 0,98 uz 95% interval pouzdanosti 0,94-1,00). Rezultati usporedbe kvantifikacije DNA EBV-a pokazali su statistički značajnu razliku u viremijama (medijan 49 400 vs. 128 000 kopija DNA EBV-a, p<0.001). Rezultati ovog istraživanja pokazuju da se praćenje kinetike replikacije virusa EBV u biomedicinskim istraživanjima obvezno mora temeljiti na primjeni identičnog standardiziranog molekularnog testa u kompletnom istraživanju te da rezultati između dostupnih standardiziranih testova nisu u potpunosti usporedivi.
Abstract (english)
Epstein-Barr virus (EBV) is an oncogenic virus that belongs to the family Orthoherpesviridae, genus Lymphocryptovirus, and has a double-stranded DNA genome that consists of 172 kb. The main cellular targets for viral replication via latent and lytic cycles include epithelial cells, B-lymphocytes, T-lymphocytes and NK-cells. The aim of this study was to compare the number of EBV DNA copies in peripheral blood samples determined by using two standardized real-time quantitative PCR test and to determine whether the results of the viral replication kinetics assessment are comparable and applicable in biological and biomedical research. EBV DNA detection in 100 peripheral blood samples by using 2 real-time PCR assays (LightMix LightCycler 2.0 and LightMix LightCycler 480 II with adapted protocol) showed concordant results in 99 of 100 samples (Kohen's kappa coefficient 0,98, 95% confidence interval 0,94-1,00). Comparison of EBV DNA quantification results obtained by the two assays has shown a statistically significant difference in measured viremias (median 49 400 vs. 128 000 EBV DNA copies/ml, p< 0.001). The results of this study have shown that monitoring of EBV quantification in biomedical research needs to be based on consistent use of identical molecular assays and that the results of standardised assays available today are not completely comparable.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:180479
ProjectNumber: IP-2020-02-8961
Title: Molekularna raznolikost virusa Epstein-Barr
Acronym: MRVEB
Leader: Snježana Židovec Lepej
Jurisdiction: Croatia
Funder: HRZZ
Funding stream: Istraživački projekti 2020.g
Study programmeTitle: Molecular Biology-University graduate programme
Study programme type: university
Study level: graduate
Academic / professional title: sveučilišni/a magistar/magistra molekularne biologije (sveučilišni/a magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2024-03-08 12:10:51