Comparative evaluation of PCR-based methods for the assessment of T cell clonality in the diagnosis of T cell lymphoma

Cairns, Suzanne M.; Taylor, Jeremy; Gould, Paul; Spagnolo, Dominic V.

doi:10.1080/003130202760120463

Cited by 12 publications

(10 citation statements)

References 14 publications

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“…1 Even though TCR-␥ comprises only 11 functional V regions, several different PCR strategies are in use. 13,14 We rigorously compared the recently described BIOMED-2 TCR-␥ assay, which targets two different TCR-␥ V gene segments (V1-8/V10 and V9/V11) in two separate reactions (multi-V strategy; Figure 1A) 15 against a previously described TCR-␥ laboratory-developed assay, which has been cited more than 100 times (ISI Web of knowledge citation index; Thompson Reuters, Philadelphia, PA), which targets a single TCR-␥ V gene segment (V1-8, V9, V10, V11) in each of four individual PCR reactions (mono-V strategy; Figure 1A). 16 In a sense, both TCR-␥ PCR strategies can be considered variants of multiplex strategies because they combine one or two V primers with multiple J primers in each PCR reaction: 15,16 the multi-V strategy adds the JP1/JP2 and J1/J2 primers 15 and the mono-V strategy adds the JP1/ JP2, J1/J2 and JP primers 16 ( Figure 1B).…”

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confidence: 99%

Comparison of BIOMED-2 Versus Laboratory-Developed Polymerase Chain Reaction Assays for Detecting T-Cell Receptor-γ Gene Rearrangements

Patel

Pan

Wang

et al. 2010

The Journal of Molecular Diagnostics

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Detecting clonal T-cell receptor (TCR)-␥ gene rearrangements (GRs) is an important adjunct test fordiagnosing T-cell lymphoma. We compared a recently described assay (BIOMED-2 protocol) , which targets multiple variable (V) gene segments in two polymerase chain reaction (PCR) reactions (multi-V) , with a frequently referenced assay that targets a single V gene segment in four separate PCR reactions (mono-V). A total of 144 consecutive clinical DNA samples were prospectively tested for T-cell clonality by PCR using laboratory-developed mono-V and commercial multi-V primer sets for TCR-␥ GR. The combination of TCR-␤ , mono-V TCR-␥ and multi-V TCR-␥ detected more clonal cases (68/144 , 47%) than any individual PCR assay. We detected clonal TCR-␤ GR in 47/68 (69%) cases. Using either mono-V or multi-V TCR-␥ primers , the sensitivities for detecting clonality were 52/68 (76%) or 51/68 (75%). Using both mono-V and multi-V TCR-␥ primers improved the sensitivity for detecting clonality , 60/68 (88%). Combining either mono-V or multi-V TCR-␥ primers with TCR-␤ primers also improved the sensitivity , 64/68 (94%). Significantly , TCR-␥ V11 GRs could only be detected using the mono-V-PCR primers. We conclude that using more than one T-cell PCR assay can enhance the overall sensitivity for detecting T-cell clonality.

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confidence: 99%

Comparison of BIOMED-2 Versus Laboratory-Developed Polymerase Chain Reaction Assays for Detecting T-Cell Receptor-γ Gene Rearrangements

Patel

Pan

Wang

et al. 2010

The Journal of Molecular Diagnostics

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“…68,79,[91][92][93][94]97,133,140,145 (iv) Qualitative sensitivity As in the assessment of IgH gene rearrangements, there is a wide range in the reported frequency of clonal detection (y60% to virtually 100%), affected by the casemix, nature of the PCR assay employed, primer selection and sensitivity of the detection system. 39,47,57,76,77,100,146 Using multiple primer combinations, which will detect all possible TCRc gene rearrangements, and routine PAGE, clonal detection rates of 80-90% may be achieved. 17,47,48,98,132,139,146 This increases to w90% and approaches 100% if high-resolution complex gels 57,76,77,80 or automated fluorescent DNA fragment analysis 68,79,91,92,94,145 are employed.…”

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confidence: 99%

“…17,47,57,76,77,98,124,128,130,[137][138][139][140] (iii) PCR product detection systems Non-denaturing polyacrylamide gels of sufficient resolution are the minimum requirement to ensure adequate separation of products on the basis of their size, given that there is relatively limited junctional diversity generated during TCRc gene rearrangement. More complex denaturing gels afford greater product resolution.…”

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confidence: 99%

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The role of molecular studies in lymphoma diagnosis: a review

Spagnolo

Ellis²,

Juneja

et al. 2004

Pathology

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“…3 Therefore, complementary methods such as the assessment of T-cell clonality are generally required. Molecular techniques aiming at identifying monoclonal rearrangements of the gene loci of the T-cell receptor (TCR)g or chains are the gold standard, [4][5][6] revealing the presence of a T-cell clone in 60-100% of peripheral T-cell lymphomas [7][8][9][10] with good specificity, albeit not 100%. 10 Although very useful in routine diagnosis, molecular assays are not quantitative, and do not allow the immunological characterization of the T-cell clone, which is a requisite for improving our knowledge of the malignant T cells involved in peripheral T-cell lymphomas.…”

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confidence: 99%

Accurate detection of the tumor clone in peripheral T-cell lymphoma biopsies by flow cytometric analysis of TCR-Vβ repertoire

et al. 2012

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Multiparametric flow cytometry has proven to be a powerful method for detection and immunophenotypic characterization of clonal subsets, particularly in lymphoproliferative disorders of the B-cell lineage. Although in theory promising, this approach has not been comparably fulfilled in mature T-cell malignancies. Specifically, the T-cell receptor-Vb repertoire analysis in blood can provide strong evidence of clonality, particularly when a single expanded V family is detected. The purpose of this study was to determine the relevance of this approach when applied to biopsies, at the site of tumor involvement. To this end, 30 peripheral T-cell lymphoma and 94 control biopsies were prospectively studied. Vb expansions were commonly detected within CD4 þ or CD8 þ T cells (97% of peripheral T-cell lymphoma and 54% of non-peripheral T-cell lymphoma cases); thus, not differentiating malignant from reactive processes. Interestingly, we demonstrated that using a standardized evaluation, the detection of a high Vb expansion was closely associated with diagnosis of peripheral T-cell lymphoma, with remarkable specificity (98%) and sensitivity (90%). This approach also identified eight cases of peripheral T-cell lymphoma that were not detectable by other forms of immunophenotyping. Moreover, focusing Vb expression analysis to T-cell subsets with aberrant immunophenotypes, we demonstrated that the T-cell clone might be heterogeneous with regard to surface CD7 or CD10 expression (4/11 cases), providing indication on 'phenotypic plasticity'. Finally, among the wide variety of Vb families, the occurrence of a Vb17 expansion in five cases was striking. To our knowledge, this is the first report demonstrating the power of T-cell receptor-Vb repertoire analysis by flow cytometry in biopsies as a basis for peripheral T-cell lymphoma diagnosis and precise T-cell clone identification and characterization.

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Comparative evaluation of PCR-based methods for the assessment of T cell clonality in the diagnosis of T cell lymphoma

Cited by 12 publications

References 14 publications

Comparison of BIOMED-2 Versus Laboratory-Developed Polymerase Chain Reaction Assays for Detecting T-Cell Receptor-γ Gene Rearrangements

Comparison of BIOMED-2 Versus Laboratory-Developed Polymerase Chain Reaction Assays for Detecting T-Cell Receptor-γ Gene Rearrangements

The role of molecular studies in lymphoma diagnosis: a review

Accurate detection of the tumor clone in peripheral T-cell lymphoma biopsies by flow cytometric analysis of TCR-Vβ repertoire

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