Optimizacija panela protiteles za diagnostiko B - celičnih limfomov

Kos, Tjaša (author), Kloboves Prevodnik, Veronika (mentor)

AI Imunofenotipizacija s pretočnim citometrom (IPC) je že dolgo uveljavljena metoda za diagnozo B-celičnih limfomov in klasifikacijo le teh. Metoda se z vedno zmoglivejšimi pretočnimi citometri hitro razvija. Prav tako je hiter tudi razvoj označevalcev. Vedno več laboratorijev po svetu uporablja 8-10 barvne osnovne panele protiteles (Pt) za diagnostiko B-celičnih limfomov (BCL). Z našo študijo smo želeli pokazati, kako zasnova in testiranje novega panela poteka, ter pokazati prednosti novega 10-barvnega osnovnega panela. Najprej smo testirali delovanje Pt, ki smo jih želeli uporabiti v novem panelu. Osem Pt smo preverili že predhodno v sklopu testiranja 14-barvnega panela za nizkocelularne vzorce, 2 Pt pa smo testirali naknadno. Prav tako smo naknadno testirali tudi delovanje Pt CD23-APC-R700, s katerim smo nadomestili Pt CD23- APC-AF700, ki ni delalo dobro. Po testiranju delovanja Pt, smo naredili nastavitve pretočnega citometra, kompenzacijo in titracijo Pt. Na koncu smo nov 10-barvni osnovni panel testirali na 48 vzorcih. Njegovo delovanje smo preveriti na 10 vzorcih reaktivnih limfocitnih proliferacij (RLP) in na 38 vzorcih BCL. Vključili smo najpogostejše BCL, ki jih dnevno analiziramo v laboratoriju: folikularni limfom (FL), limfom plaščnih celic (LPC), kronično limfatično levkemijo (KLL), difuzni velikocelični limfom B (DVCLB), limfom marginalne cone (LMC). Nov 10-barvni osnovni panel smo testirali tudi na 2 primerih kompozitnih limfomov. Z novim panelom smo prišli do enakih rezultatov, kot z rutinskim 4-barvnim osnovnim panelom, s čimer smo potrdili, da je nov 10-barvni osnovni panel primeren za uporabo v vsakodnevni rutinski diagnostiki BCL.

• Language: Slovenian
• Keywords: panel protiteles, pretočna citometrija, limfom
• Work type: Master's thesis/paper
• Organization: BF - Biotechnical Faculty
• Year: 2018
• PID: 20.500.12556/RUL-105729
• COBISS.SI-ID: 4927823
• Publication date in RUL: 08.12.2018

Secondary language

• Language: Unknown
• Title: Optimisation of the panel of antibodies for the B-cell lymphomas diagnosis

Abstract

Flow cytometric immunophenotyping (FCI) is a long-established method for diagnosis and classification of B-cell lymphomas. The method is rapidly evolving with better-performing flow cytometers. The development of markers is also fast. More and more laboratories around the world use 8-10 colored basic antibody (Ab) panels for the diagnosis of B-cell lymphomas (BCL). With our study, we wanted to show how the design and testing of the new panel is taking place and show the benefits of the new 10-color panel. We first tested the Ab performance we wanted to use in the new panel. Eight Ab was checked previously in the framework of testing the 14-color panel for low-cellular samples, and 2 Ab was subsequently tested. We also subsequently tested the Ab CD23-APC-R700 performance, because of Ab CD23-APC-AF700, which did not work well. After testing the Ab performance, we made the flow cytometer settings, compensation and Ab titration. In the end, a new 10-color panel was tested on 48 samples. Its performance was checked on 10 samples of reactive lymphocytic proliferation (RLP) and on 38 BCL samples. We included the most common BCLs, which are analyzed daily in the laboratory: follicular lymphoma (FL), mantle cell lymphoma (MCL), chronic lymphatic leukemia (CLL), diffuse large B cell lymphoma (DLBCL), marginal zone lymphoma (MZL). A new 10-color panel was also tested on 2 cases of composite lymphomas. With the new panel, we got the same results as with the routine 4-color panel, confirming that the new 10-color base panel is suitable for use in day-to-day BCL routine diagnostics.

• Keywords: antibody panel, flow cytometry, lymphoma