Detection of t(14;18) by PCR of IgH/BCL2 fusion gene in follicular lymphoma from archived cytological smears

Štoos-Veić, Tajana and Livun, Ana and Ajduković, Radmila and Pejša, Vlatko and Jakšić, Ozren and Kušec, Rajko (2010) Detection of t(14;18) by PCR of IgH/BCL2 fusion gene in follicular lymphoma from archived cytological smears. Collegium Antropologicum, 34 (2). pp. 425-9. ISSN 0350-6134

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Abstract

According to WHO classification follicular lymphoma (FL) is a neoplasm composed of follicle centre (germinal centre) B-cells, which usually has at least a partially follicular pattern. Bone marrow (BM) infiltration by lymphoma occurs in 40-70% of cases at the time of diagnosis. The characteristic chromosomal translocation of follicular lymphoma is t(14;18)(q32;q21) with transposition of BCL2 oncogene to the regulatory region of immunoglobulin heavy chain gene IgH. Aim of this study was to determine the frequency of PCR detection of IgH/BCL2 in DNA samples isolated from archival cytological slides of lymph node aspirates, bone marrow and/or peripheral blood (PB) obtained from patients with histologically confirmed follicular lymphoma using primers and protocol proposed by BIOMED-2 consortium. We also compared molecular with cytomorphological findings in bone marrow/peripheral blood and tested this method of detection of IgH/BCL2 molecular marker in monitoring minimal residual disease (MRD) in routine clinical setting. DNA was successfully isolated from all archival cytological slides obtained by fine needle aspiration of lymph nodes as well as from 75% of smears of bone marrow aspirates from 19 patients. Fusion oncogene was detected in 10 of 19 patients (52%). For patients with PCR IgH/BCL2 positive lymph nodes, molecular test found BM infiltration in 5 cases (83%), while cytomorphology detected infiltration in three of eight cases (37%) available for comparison. May-Grünwald-Giemsa stained cytological smears can be used for PCR-based ancillary methods and the rate of detection of IgH/BCL2 rearrangement is similar to results reported for paraffin-embedded tissues. For patients with detectable baseline molecular marker, PCR is a highly suitable method for detection of bone marrow involvement and monitoring MRD.

Abstract in Croatian

PCR ZA OTKRIVANJE IGH/BCL2 PREUREDBE S ARHIVSKIH CITOLOŠKIH RAZMAZA PUNKTATA FOLIKULARNIH LIMFOMA ----- Prema klasifikaciji Svjetske zdravstvene organizacije, folikularni limfomi su tumori stanica centra folikula koji bar djelomice zadržavaju folikularni način rasta. Infiltracija koštane srži nalazi se u 40–70% slučajeva u vrijeme postavljanja dijagnoze. Karakteristična kromosomska translokacija folikularnih limfoma je t(14;18)(q32;q21) koja premješta BCL2 onkogen pod kontrolu promotorske regije za teške lance imunoglobulina IgH na kromosomu 14. Cilj ovog rada bio je odrediti učestalost translokacije t(14;18) molekularnom analizom temeljenom na polimeraza lančanoj reakciji (PCR) u uzorcima DNA izoliranim s arhivskih citoloških stakala koristeći standardizirani analitički protokol Europske radne grupe BIOMED-2, usporediti molekularne i citomorfološke nalaze te ispitati vrijednost metode za praćenje minimalne ostatne bolesti (MRD) u svakodnevnim uvjetima. Izolacija DNA je uspjela sa svih citoloških razmaza punktata limfnih čvorova kao i u 75% razmaza punktata koštane srži, a PCR-om je otkrivena IgH/BCL2 preuredba u 10 od 19 (52%) bolesnika. U bolesnika s pozitivnim molekularnim testom u limfnom čvoru, PCR-om je pronađena infiltracija koštane srži limfomom u 5 bolesnika (83%), dok je citološka analiza infiltraciju našla u 3 od osam uzoraka dostupnih za usporedbu. Citološki razmazi bojani May-Grünwald Giemsa metodom (MGG) dobar su izvor DNA za metode molekularne analize temeljene na lančanoj reakciji polimeraze. Učestalost otkrivanja IgH/BCL2 preuredbe odgovara rezultatima studija rađenih na histološkim uzorcima. U bolesnika u kojih je pronađena translokacija u dijagnostičkim uzorcima, PCR je pogodna metoda za otkrivanje infiltracije koštane srži limfomom te za praćenje minimalne ostatne bolesti.

Item Type: Article
Departments: Katedra za internu medicinu
Depositing User: Marijan Šember
Status: Published
Creators:
CreatorsEmail
Štoos-Veić, TajanaUNSPECIFIED
Livun, AnaUNSPECIFIED
Ajduković, RadmilaUNSPECIFIED
Pejša, VlatkoUNSPECIFIED
Jakšić, OzrenUNSPECIFIED
Kušec, RajkoUNSPECIFIED
Date: June 2010
Date Deposited: 01 Sep 2010
Last Modified: 23 Sep 2011 16:11
Subjects: /
Related URLs:
    URI: http://medlib.mef.hr/id/eprint/847

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