20.500.12556/RUL-170810 Implementacija platforme Dako Omnis za določanje limfomskih označevalcev na citoloških vzorcih Implementation of the Dako Omnis platform for assessing lymphoma markers on cytology samples Diagnostika vseh vrst rakavih obolenj, med njimi tudi limfomov, je področje, ki se nenehno razvija. Za natančno določanje limfomskih označevalcev v citoloških in histoloških vzorcih se danes uporabljajo najrazličnejše metode, ki temeljijo na histologiji, imunohistokemiji/imunocitokemiji (IHK/ICK), pretočni citometriji in molekularnih metodah. Na trg prihajajo vedno nove platforme, ki bi lahko izboljšale in pohitrile diagnostiko limfomov. Ena izmed takih platform je IHK/ICK platforma Dako Omnis. Namen magistrske naloge je bil preveriti učinkovitost in zanesljivost popolnoma avtomatizirane platforme Dako Omnis za ICK barvanje limfomskih označevalcev na citoloških vzorcih ter rezultate primerjati avtomatizirano platformo BenchMark Ultra, ki se na Onkološkem inštitutu Ljubljana (OIL) uporablja v vsakodnevni, rutinski diagnostiki. Na platformi Dako Omnis smo optimizirali in validirali protokole za določanje limfomskih označevalcev CD45, CD3, CD20, CD5, CD10, CD30, CD15, К, λ, Ki-67 in PAX5. Ker je nastanek številnih limfomov povezan z okužbo z virusom Epstein barr (EBV), smo dodatno uvedli nov protokol za določanje prisotnosti EBV v citoloških vzorcih. Ugotovili smo, da platforma Dako Omnis omogoča izvedbo ICK barvanja, vendar rezultati pri večini limfomskih označevalcev niso bili popolnoma primerljivi z rezultati ICK barvanja na platformi BenchMark Ultra in z rezultati pretočne citometrije. Rezultati kažejo, da ima platforma Dako Omnis potencial, a so za rutinsko uporabo na citoloških vzorcih potrebne nadaljnje izboljšave in standardizacija. Predvsem bi bilo ICK barvanje potrebno optimizirati in validati na celičnih blokih in ICK barvanje na cistospinih izvajati še naprej na platformi BenchMark Ultra. Diagnostics of various types of cancer, including lymphomas, is a continuously evolving field. A wide array of methods is currently used to accurately determine lymphoma markers in cytological and histological samples, including histology, immunohistochemistry/ immunocytochemistry (IHC/ICC), flow cytometry, and molecular techniques. New platforms are regularly introduced to the market with the aim of improving and accelerating lymphoma diagnostics; one such IHC/ICC platform is Dako Omnis. The purpose of this master’s thesis was to evaluate the efficiency and reliability of the fully automated Dako Omnis platform for ICC staining of lymphoma markers in cytological samples and to compare the results with those obtained using the automated BenchMark Ultra platform, which is routinely used at the Institute of Oncology Ljubljana. On the Dako Omnis platform, protocols were optimized and validated for lymphoma markers CD45, CD3, CD20, CD5, CD10, CD30, CD15, κ, λ, Ki-67, and PAX5. Since the development of many lymphomas is associated with Epstein-Barr virus (EBV) infection, a new protocol was also introduced for detecting EBV in cytological samples. The study found that the Dako Omnis platform is capable of performing ICC staining, however, for most lymphoma markers, the results were not entirely comparable with those obtained using the BenchMark Ultra platform or flow cytometry. The findings suggest that while the Dako Omnis platform shows potential, further improvements and standardization are necessary before it can be used routinely on cytological samples. Specifically, ICC staining should be optimized and validated on cell blocks, while ICC staining on cytospins should continue to be carried out using the BenchMark Ultra platform. Limfom imunocitokemija pretočna citometrija platforma Dako Omnis detekcisjki sistem Envision FLEX Lymphoma immunocytochemistry flow cytometry Dako Omnis platform Envision FLEX detection system true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-07-17 07:15:30 2025-07-17 07:15:38 2025-07-19 03:40:49 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 257828 242874115 49474.pdf 49474.pdf 1 95726F1EED63492A7A8ACEC25632AEC6 19376029b41551a976f72250eb668af5c3e8fe797260863d1fc46fd5b87d222a 2204014d-62cb-11f0-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=213019 Biotehniška fakulteta 0 0 0