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Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections

Objective: Follicular lymphoma (FL) is one of the most common lymphomas, and is characterized by t(14;18)(q32;q21) in more than 80% of patients. The aim of this study was to determine the rate of t(14;18) positivity based onthe detection of mbr or mcr in paraffin-embedded tissue samples. Material an...

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Autores principales: Selvi, Nur, Kosova, Buket, Hekimgil, Mine, Gündüz, Cumhur, Kaymaz, Burçin Tezcanlı, Karaca, Emin, Saydam, Güray, Tombuloğlu, Murat, Büyükkeçeci, Filiz, Çağırgan, Seçkin, Ertan, Yeşim, Topçuoğlu, Nejat
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Galenos Publishing 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3986950/
https://www.ncbi.nlm.nih.gov/pubmed/24744643
http://dx.doi.org/10.5505/tjh.2012.93898
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author Selvi, Nur
Kosova, Buket
Hekimgil, Mine
Gündüz, Cumhur
Kaymaz, Burçin Tezcanlı
Karaca, Emin
Saydam, Güray
Tombuloğlu, Murat
Büyükkeçeci, Filiz
Çağırgan, Seçkin
Ertan, Yeşim
Topçuoğlu, Nejat
author_facet Selvi, Nur
Kosova, Buket
Hekimgil, Mine
Gündüz, Cumhur
Kaymaz, Burçin Tezcanlı
Karaca, Emin
Saydam, Güray
Tombuloğlu, Murat
Büyükkeçeci, Filiz
Çağırgan, Seçkin
Ertan, Yeşim
Topçuoğlu, Nejat
author_sort Selvi, Nur
collection PubMed
description Objective: Follicular lymphoma (FL) is one of the most common lymphomas, and is characterized by t(14;18)(q32;q21) in more than 80% of patients. The aim of this study was to determine the rate of t(14;18) positivity based onthe detection of mbr or mcr in paraffin-embedded tissue samples. Material and Methods: The study included 32 paraffin-embedded tissue samples collected from 32 consecutive FL patients that were diagnosed and followed-up at our hospital between 1999 and 2006. The MBR breakpoint wasidentified based on real-time PCR using a LightCycler v.2.0 t(14;18) Quantification Kit (MBR), multiplex PCR, and seminestedPCR. To identify the mcr breakpoint, real-time PCR was performed using specific primers and the FastStart DNAMaster SYBR Green I Kit. To detect t(14;18) via fluorescence in situ hybridization (FISH) nuclei from paraffin-embeddedtissue sections were extracted and used together with LSI IgH (immunoglobulin heavy chain) (spectrum green)/bcl-2(B-cell leukemia-lymphoma 2) (spectrum orange) probes. Results: The DNA and nuclei isolation success rate for B5 formalin-fixed, paraffin-embedded tissue sections (n = 12)was 42% and 33%, respectively, versus 95% and 60%, respectively, for 20 tissue sections fixed in formalin only. In all,24 paraffin-embedded tissue sections were analyzed and mbr positivity was observed in the DNA of 82.14% via seminested PCR, in 53.57% via multiplex PCR, and in 28.57% via real-time PCR. We did not detect mcr rearrangementin any of the samples. In all, 15 of 16 patients (93.75%) whose nuclei were successfully isolated were observed to bet(14;18) positive via the FISH method. Conclusion: Semi-nested PCR and FISH facilitated the genetic characterization of FL tumors. As such, FISH and PCR complement each other and are both essential for detecting t(14;18) translocation.
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spelling pubmed-39869502014-04-17 Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections Selvi, Nur Kosova, Buket Hekimgil, Mine Gündüz, Cumhur Kaymaz, Burçin Tezcanlı Karaca, Emin Saydam, Güray Tombuloğlu, Murat Büyükkeçeci, Filiz Çağırgan, Seçkin Ertan, Yeşim Topçuoğlu, Nejat Turk J Haematol Research Article Objective: Follicular lymphoma (FL) is one of the most common lymphomas, and is characterized by t(14;18)(q32;q21) in more than 80% of patients. The aim of this study was to determine the rate of t(14;18) positivity based onthe detection of mbr or mcr in paraffin-embedded tissue samples. Material and Methods: The study included 32 paraffin-embedded tissue samples collected from 32 consecutive FL patients that were diagnosed and followed-up at our hospital between 1999 and 2006. The MBR breakpoint wasidentified based on real-time PCR using a LightCycler v.2.0 t(14;18) Quantification Kit (MBR), multiplex PCR, and seminestedPCR. To identify the mcr breakpoint, real-time PCR was performed using specific primers and the FastStart DNAMaster SYBR Green I Kit. To detect t(14;18) via fluorescence in situ hybridization (FISH) nuclei from paraffin-embeddedtissue sections were extracted and used together with LSI IgH (immunoglobulin heavy chain) (spectrum green)/bcl-2(B-cell leukemia-lymphoma 2) (spectrum orange) probes. Results: The DNA and nuclei isolation success rate for B5 formalin-fixed, paraffin-embedded tissue sections (n = 12)was 42% and 33%, respectively, versus 95% and 60%, respectively, for 20 tissue sections fixed in formalin only. In all,24 paraffin-embedded tissue sections were analyzed and mbr positivity was observed in the DNA of 82.14% via seminested PCR, in 53.57% via multiplex PCR, and in 28.57% via real-time PCR. We did not detect mcr rearrangementin any of the samples. In all, 15 of 16 patients (93.75%) whose nuclei were successfully isolated were observed to bet(14;18) positive via the FISH method. Conclusion: Semi-nested PCR and FISH facilitated the genetic characterization of FL tumors. As such, FISH and PCR complement each other and are both essential for detecting t(14;18) translocation. Galenos Publishing 2012-06 2012-06-15 /pmc/articles/PMC3986950/ /pubmed/24744643 http://dx.doi.org/10.5505/tjh.2012.93898 Text en © Turkish Journal of Hematology, Published by Galenos Publishing. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Selvi, Nur
Kosova, Buket
Hekimgil, Mine
Gündüz, Cumhur
Kaymaz, Burçin Tezcanlı
Karaca, Emin
Saydam, Güray
Tombuloğlu, Murat
Büyükkeçeci, Filiz
Çağırgan, Seçkin
Ertan, Yeşim
Topçuoğlu, Nejat
Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections
title Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections
title_full Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections
title_fullStr Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections
title_full_unstemmed Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections
title_short Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections
title_sort molecular evaluation of t(14;18)(bcl-2/igh) translocation in follicular lymphoma at diagnosis using paraffin-embedded tissue sections
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3986950/
https://www.ncbi.nlm.nih.gov/pubmed/24744643
http://dx.doi.org/10.5505/tjh.2012.93898
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