title: Real-time 5'-->3' exonuclease-based PCR assay for detection of the t(11;14)(q13;q32).
abstract: We describe the usefulness of a real-time polymerase chain reaction (PCR) assay for detection of the t(11;14)(q13;q32), most commonly present in mantle cell lymphoma (MCL). This assay is based on the 5'-->3' exonuclease activity of Taq polymerase, which cleaves an internal probe labeled with a reporter dye at its 5' end and a quencher dye at its 3' end during PCR. The real-time t(11;14) PCR assay was established using DNA from a case of MCL with the t(11;14), amplifiable using conventional PCR and primers specific for the major translocation cluster (MTC) region of the bcl-1 locus and the immunoglobulin heavy chain joining region gene (JH). The specificity was determined by analyzing DNA from 82 cases: 50 MCL, 27 other types of non-Hodgkin lymphoma (NHL), and 5 reactive lymphoid proliferations. The real-time t(11;14) PCR results were correlated with data obtained by a conventional PCR assay. By using the real-time assay, bcl-1 MTC/JH DNA fusion sequences were detected in 25 of 50 MCLs but not in other NHLs or reactive lymphoid proliferations. Concordance between real-time and conventional PCR methods for MCL was 96% and for all samples was 98%. The results demonstrate that this real-time PCR method to detect bcl-1 MTC/JH DNA fusion sequences is specific and reliable. In addition, the results are available immediately following amplification, without standard post-PCR manipulations.
authors.affiliation: Department of Pathology, University of Texas MD, Anderson Cancer Center, Houston 77030, USA.
authors.forename: R
authors.initials: R
authors.lastname: Luthra
authors.forename: A H
authors.initials: AH
authors.lastname: Sarris
authors.forename: S
authors.initials: S
authors.lastname: Hai
authors.forename: A V
authors.initials: AV
authors.lastname: Paladugu
authors.forename: J E
authors.initials: JE
authors.lastname: Romaguera
authors.forename: F F
authors.initials: FF
authors.lastname: Cabanillas
authors.forename: L J
authors.initials: LJ
authors.lastname: Medeiros
mesh_terms: D002880:Chromosomes, Human, Pair 11; D002883:Chromosomes, Human, Pair 14; D003199:Computer Systems; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007133:Immunoglobulin Joining Region / Q000235:genetics; D008228:Lymphoma, Non-Hodgkin / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D019914:Taq Polymerase / Q000378:metabolism; D014178:Translocation, Genetic
chemical_list: D007143:Immunoglobulin Heavy Chains; D007133:Immunoglobulin Joining Region; D019914:Taq Polymerase; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V