Molekularne metode prikladne za praćenje kimerizma nakon transplantacije krvotvornih matičnih stanica

master's thesis

• Title (english): Molecular methods of choice for chimerism monitoring after hematopoietic stem cell transplantation
• Author: Jana Jančin
• Mentor: Zorana Grubić (mentor)
• Committee member: Zorana Grubić (predsjednik povjerenstva)
• Committee member: Ana Galov (član povjerenstva)
• Committee member: Petra Korać (član povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2022-09-27, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Uspjeh transplantacije krvotvornih matičnih stanica (TKM) procjenjuje se analizom statusa
kimerizma odnosno, udjela primateljevih stanica u uzorku bolesnika nakon TKMS. Cilj
istraživanja bio je usporediti dvije molekularne metode u praćenju kimerizma: metodu umnažanja
kratkih uzastopnih ponavljanja DNA lančanom reakcijom (PCR-STR) i metodu lančane reakcije
polimerazom u stvarnom vremenu (qPCR) u svrhu određivanja pouzdanosti, osjetljivosti i
preciznosti metoda ovisno o omjeru primateljevih i davateljevih stanica u uzorku bolesnika nakon
TKMS. Istraživanje je obuhvatilo 12 parova primatelj-davatelj te sveukupno 42 uzorka kimera u
kojima je udio primateljevih stanica bio u rasponu od 10-90%. Od 21 analiziranog lokusa STR,
najinformativnijima su se pokazali lokusi D13S797, D18S976 i D21S1442, dok je među 30 biljega
qPCR za daljnu analizu su najčešće upotrebljavani biljezi KMR040 i KMR050. Raspon dobivenih
vrijednosti za udio primateljevih stanica su u prosjeku bio je manji za lokuse STR nego raspon
dobiven analizom biljega qPCR. Istraživanje je pokazalo da je metoda qPCR manje pouzdana od
metode PCR-STR u određivanju udjela primateljevih stanica kod bolesnika s >10% udjela vlastitih
stanica u uzorku nakon TKMS. Zaključno, metoda PCR-STR je bolji izbor za praćenje kimerizma
kod bolesnika s visokim udjelom primateljevih stanica.

Abstract (english)

Success of hematopoietic stem cell transplantation (HSCT) is evaluated by chimerism analysis,
which is based on determination of recipient and donor cells ratio in patient's sample after HSCT.
Purpose of this study was to compare two molecular methods in chimerism monitoring: Short
Tandem Repeat-Polymerase Chain Reaction (STR-PCR) and Real time-Polymerase Chain
Reaction (qPCR), in order to determine their reliability, sensitivity and accuracy depending on
ratio of recipients and donors cells in recipient’s sample after HSCT. This study included 12
recipient-donor pairs and altogether 42 post-HSCT samples in which recipient’s cell ratio was in
the range of 10-90%. Out of 12 analysed STR loci the most informative ones were D13S797,
D18S976 and D21S1442, while among 30 investigated qPCR markers, KMR040 and KMR050
were most frequently used for chimerism analysis. Range of values obtained for percentage of
recipient cells was lower for STR loci in comparison to the one obtained by analysis of qPCR
markers. The study showed that qPCR is less reliable then STR-PCR for chimerism monitoring
for patients with >10% of their own cells in post HSCT sample. In conclusion, PCR-STR method
is a better choice for chimerism status monitoring of those patients with a higher percentage of
recipient cells.

• Keywords: PCR-STR; qPCR; informativni lokusi
• Keywords (english): PCR-STR; qPCR; informative loci
• Language: croatian
• URN:NBN: urn:nbn:hr:217:524410
• Study programme: Title: Experimental Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2023-09-27
• Created on: 2022-10-21 10:18:54