Combined molecular diagnosis of b-cell lymphomas with t(11;14)(q13;q32) or t(14;18)(q32;q21) using multiplex- and long distance inverse-polymerase chain reaction

Luís Vieira, Ana Martinho, Ofélia Antunes, Elizabeth Silva, Ana P. Ambrósio, Maria C. Geraldes, Rute Nascimento, Cĝndido Silva, José M. Pereira, Esmeraldina C. Júnior, Peter Jordan

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Translocations t(14;18)(q32;q21) and t(11;14)(q13;q32) are recurrent findings in follicular lymphoma (FL) and mantle cell lymphoma (MCL), respectively. However, the molecular counterparts of these translocations can only be detected in up to 75% of FL and 50% of MCL cases using routine techniques. To improve the efficiency of detection, we first devised a single-tube multiplex-polymerase chain reaction (PCR) assay with primers located within a conserved immunoglobulin heavy chain (IGH) sequence and 5′ to the main breakpoint cluster regions of BCL2 and CCND1. Using this assay in 17 FL and 11 MCL diagnostic DNA samples, we readily identified a BCL2-IGH fusion in 65% of FL patients and a CCND1-IGH fusion in 55% of MCL patients. In the remaining cases, we used long distance inverse-PCR to detect BCL2-IGH and CCND1-IGH fusion genes with different BCL2 and CCND1 breakpoint locations. We found additional translocations in 3 patients (17%) with FL and in 4 patients (36%) with MCL. Taken together, we show that multiplex-PCR combined with long distance inverse-PCR detected a t(14;18) in 82% of FL patients and a t(11;14) in 91% of MCL patients, demonstrating that this 2-step protocol is an effective approach for molecular detection of t(11;14) and t(14;18) in B-cell lymphomas.

Original languageEnglish
Pages (from-to)73-81
Number of pages9
JournalDiagnostic Molecular Pathology
Volume17
Issue number2
DOIs
Publication statusPublished - 1 Jun 2008

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Mantle-Cell Lymphoma
Follicular Lymphoma
Immunoglobulin Heavy Chains
Lymphoma
Polymerase Chain Reaction
Multiplex Polymerase Chain Reaction
Immunoglobulin Heavy Chain Genes
B-Cell Lymphoma
DNA

Keywords

  • BCL2
  • CCND1
  • IGH
  • Long distance inverse-PCR
  • Multiplex-PCR

Cite this

Vieira, Luís ; Martinho, Ana ; Antunes, Ofélia ; Silva, Elizabeth ; Ambrósio, Ana P. ; Geraldes, Maria C. ; Nascimento, Rute ; Silva, Cĝndido ; Pereira, José M. ; Júnior, Esmeraldina C. ; Jordan, Peter. / Combined molecular diagnosis of b-cell lymphomas with t(11;14)(q13;q32) or t(14;18)(q32;q21) using multiplex- and long distance inverse-polymerase chain reaction. In: Diagnostic Molecular Pathology. 2008 ; Vol. 17, No. 2. pp. 73-81.
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title = "Combined molecular diagnosis of b-cell lymphomas with t(11;14)(q13;q32) or t(14;18)(q32;q21) using multiplex- and long distance inverse-polymerase chain reaction",
abstract = "Translocations t(14;18)(q32;q21) and t(11;14)(q13;q32) are recurrent findings in follicular lymphoma (FL) and mantle cell lymphoma (MCL), respectively. However, the molecular counterparts of these translocations can only be detected in up to 75{\%} of FL and 50{\%} of MCL cases using routine techniques. To improve the efficiency of detection, we first devised a single-tube multiplex-polymerase chain reaction (PCR) assay with primers located within a conserved immunoglobulin heavy chain (IGH) sequence and 5′ to the main breakpoint cluster regions of BCL2 and CCND1. Using this assay in 17 FL and 11 MCL diagnostic DNA samples, we readily identified a BCL2-IGH fusion in 65{\%} of FL patients and a CCND1-IGH fusion in 55{\%} of MCL patients. In the remaining cases, we used long distance inverse-PCR to detect BCL2-IGH and CCND1-IGH fusion genes with different BCL2 and CCND1 breakpoint locations. We found additional translocations in 3 patients (17{\%}) with FL and in 4 patients (36{\%}) with MCL. Taken together, we show that multiplex-PCR combined with long distance inverse-PCR detected a t(14;18) in 82{\%} of FL patients and a t(11;14) in 91{\%} of MCL patients, demonstrating that this 2-step protocol is an effective approach for molecular detection of t(11;14) and t(14;18) in B-cell lymphomas.",
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Vieira, L, Martinho, A, Antunes, O, Silva, E, Ambrósio, AP, Geraldes, MC, Nascimento, R, Silva, C, Pereira, JM, Júnior, EC & Jordan, P 2008, 'Combined molecular diagnosis of b-cell lymphomas with t(11;14)(q13;q32) or t(14;18)(q32;q21) using multiplex- and long distance inverse-polymerase chain reaction', Diagnostic Molecular Pathology, vol. 17, no. 2, pp. 73-81. https://doi.org/10.1097/PDM.0b013e31814be9e0

Combined molecular diagnosis of b-cell lymphomas with t(11;14)(q13;q32) or t(14;18)(q32;q21) using multiplex- and long distance inverse-polymerase chain reaction. / Vieira, Luís; Martinho, Ana; Antunes, Ofélia; Silva, Elizabeth; Ambrósio, Ana P.; Geraldes, Maria C.; Nascimento, Rute; Silva, Cĝndido; Pereira, José M.; Júnior, Esmeraldina C.; Jordan, Peter.

In: Diagnostic Molecular Pathology, Vol. 17, No. 2, 01.06.2008, p. 73-81.

Research output: Contribution to journalArticle

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T1 - Combined molecular diagnosis of b-cell lymphomas with t(11;14)(q13;q32) or t(14;18)(q32;q21) using multiplex- and long distance inverse-polymerase chain reaction

AU - Vieira, Luís

AU - Martinho, Ana

AU - Antunes, Ofélia

AU - Silva, Elizabeth

AU - Ambrósio, Ana P.

AU - Geraldes, Maria C.

AU - Nascimento, Rute

AU - Silva, Cĝndido

AU - Pereira, José M.

AU - Júnior, Esmeraldina C.

AU - Jordan, Peter

PY - 2008/6/1

Y1 - 2008/6/1

N2 - Translocations t(14;18)(q32;q21) and t(11;14)(q13;q32) are recurrent findings in follicular lymphoma (FL) and mantle cell lymphoma (MCL), respectively. However, the molecular counterparts of these translocations can only be detected in up to 75% of FL and 50% of MCL cases using routine techniques. To improve the efficiency of detection, we first devised a single-tube multiplex-polymerase chain reaction (PCR) assay with primers located within a conserved immunoglobulin heavy chain (IGH) sequence and 5′ to the main breakpoint cluster regions of BCL2 and CCND1. Using this assay in 17 FL and 11 MCL diagnostic DNA samples, we readily identified a BCL2-IGH fusion in 65% of FL patients and a CCND1-IGH fusion in 55% of MCL patients. In the remaining cases, we used long distance inverse-PCR to detect BCL2-IGH and CCND1-IGH fusion genes with different BCL2 and CCND1 breakpoint locations. We found additional translocations in 3 patients (17%) with FL and in 4 patients (36%) with MCL. Taken together, we show that multiplex-PCR combined with long distance inverse-PCR detected a t(14;18) in 82% of FL patients and a t(11;14) in 91% of MCL patients, demonstrating that this 2-step protocol is an effective approach for molecular detection of t(11;14) and t(14;18) in B-cell lymphomas.

AB - Translocations t(14;18)(q32;q21) and t(11;14)(q13;q32) are recurrent findings in follicular lymphoma (FL) and mantle cell lymphoma (MCL), respectively. However, the molecular counterparts of these translocations can only be detected in up to 75% of FL and 50% of MCL cases using routine techniques. To improve the efficiency of detection, we first devised a single-tube multiplex-polymerase chain reaction (PCR) assay with primers located within a conserved immunoglobulin heavy chain (IGH) sequence and 5′ to the main breakpoint cluster regions of BCL2 and CCND1. Using this assay in 17 FL and 11 MCL diagnostic DNA samples, we readily identified a BCL2-IGH fusion in 65% of FL patients and a CCND1-IGH fusion in 55% of MCL patients. In the remaining cases, we used long distance inverse-PCR to detect BCL2-IGH and CCND1-IGH fusion genes with different BCL2 and CCND1 breakpoint locations. We found additional translocations in 3 patients (17%) with FL and in 4 patients (36%) with MCL. Taken together, we show that multiplex-PCR combined with long distance inverse-PCR detected a t(14;18) in 82% of FL patients and a t(11;14) in 91% of MCL patients, demonstrating that this 2-step protocol is an effective approach for molecular detection of t(11;14) and t(14;18) in B-cell lymphomas.

KW - BCL2

KW - CCND1

KW - IGH

KW - Long distance inverse-PCR

KW - Multiplex-PCR

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