title: Molecular Evaluation of t(14;18)(bcl-2/IgH) Translocation in Follicular Lymphoma at Diagnosis Using Paraffin-Embedded Tissue Sections.
abstract: 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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Ege University, School of Medicine, Medical Biology Department, İzmir, Turkey.
authors.forename: Nur
authors.initials: N
authors.lastname: Selvi
authors.affiliation: Ege University, School of Medicine, Medical Biology Department, İzmir, Turkey.
authors.forename: Buket
authors.initials: B
authors.lastname: Kosova
authors.affiliation: Ege University, School of Medicine, Pathology Department, İzmir, Turkey.
authors.forename: Mine
authors.initials: M
authors.lastname: Hekimgil
authors.affiliation: Ege University, School of Medicine, Medical Biology Department, İzmir, Turkey.
authors.forename: Cumhur
authors.initials: C
authors.lastname: Gündüz
authors.affiliation: Ege University, School of Medicine, Medical Biology Department, İzmir, Turkey.
authors.forename: Burçin Tezcanlı
authors.initials: BT
authors.lastname: Kaymaz
authors.affiliation: Ege University, School of Medicine, Medical Genetics Department, İzmir, Turkey.
authors.forename: Emin
authors.initials: E
authors.lastname: Karaca
authors.affiliation: Ege University, School of Medicine, Hematology Department, İzmir, Turkey.
authors.forename: Güray
authors.initials: G
authors.lastname: Saydam
authors.affiliation: Ege University, School of Medicine, Hematology Department, İzmir, Turkey.
authors.forename: Murat
authors.initials: M
authors.lastname: Tombuloğlu
authors.affiliation: Ege University, School of Medicine, Hematology Department, İzmir, Turkey.
authors.forename: Filiz
authors.initials: F
authors.lastname: Büyükkeçeci
authors.affiliation: Ege University, School of Medicine, Hematology Department, İzmir, Turkey.
authors.forename: Seçkin
authors.initials: S
authors.lastname: Cağırgan
authors.affiliation: Ege University, School of Medicine, Pathology Department, İzmir, Turkey.
authors.forename: Yeşim
authors.initials: Y
authors.lastname: Ertan
authors.affiliation: Ege University, School of Medicine, Medical Biology Department, İzmir, Turkey.
authors.forename: Nejat
authors.initials: N
authors.lastname: Topçuoğlu
keywords: FISH; Follicular lymphoma; Multiplex PCR; Semi-nested PCR
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