title: Sensitivity and reproducibility of conventional qualitative and quantitative PCR assays for detection of the t(14;18)(q32;q21) chromosomal translocation in biopsy material from patients with follicular lymphoma.
abstract: Follicular lymphoma (FL) is characterized by the t(14;18)(q32;q21) chromosomal translocation which can be detected by polymerase chain reaction (PCR) in approximately 70% of cases. The aim of our retrospective study was to evaluate the sensitivity and the reproducibility of both conventional qualitative and quantitative PCR assays for detection of the t(14;18)(q32;q21) chromosomal translocation in biopsy material. Fifty-seven formalin-fixed, paraffin-embedded tumor lymph node (LN) specimens from 50 patients with FL were included in the study. Qualitative PCR was performed with primer sets specific for the MBR, far3'-MBR and the mcr regions, respectively. Quantitative PCR was performed using the LightCycler instrument and the LightCycler - t(14;18) Quantification Kit (MBR). The overall detection rate of the t(14;18) in our study (52.6%) was in accordance with the literature. Of the t(14;18)-positive cases, 49.1% had breakpoints within the MBR and only 3.5% had breakpoints within the mcr. The most sensitive method was LightCycler-based PCR with a detection rate of 47.4%, followed by MBR1,2 assay (43.9%). We observed good agreement between qualitative MBR1,2 and quantitative LightCycler-based assay with a slightly higher detection rate of the quantitative method. The sensitivities of both methods were in accordance with results from other studies. Since LightCycler-based assay detects only breakpoints within the MBR, qualitative PCR should be employed in routine diagnostic settings for detection of breakpoints within the mcr and far3'-MBR regions.
authors.affiliation: Department of Molecular Diagnostics, Institute of Oncology Ljubljana, Zaloska 2, 1000 Ljubljana, Slovenia.
authors.forename: Ira
authors.initials: I
authors.lastname: Kokovic
authors.forename: Barbara Jezersek
authors.initials: BJ
authors.lastname: Novakovic
authors.forename: Snjezana Frkovic
authors.initials: SF
authors.lastname: Grazio
authors.forename: Srdjan
authors.initials: S
authors.lastname: Novakovic
mesh_terms: D001706:Biopsy; D002883:Chromosomes, Human, Pair 14 / Q000737:chemistry; D002887:Chromosomes, Human, Pair 18 / Q000737:chemistry; D005260:Female; D050438:Genes, Immunoglobulin Heavy Chain; D019254:Genes, bcl-2; D006801:Humans; D008198:Lymph Nodes / Q000473:pathology*; D008224:Lymphoma, Follicular / Q000175:diagnosis* / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling; D014178:Translocation, Genetic