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Francisco Vega, L. Jeffrey Medeiros, Dan Jones, Lynne V. Abruzzo, Raymond Lai, John Manning, Valerie Dunmire, Rajyalakshmi Luthra, A Novel Four-Color PCR Assay to Assess T-Cell Receptor Gamma Gene Rearrangements in Lymphoproliferative Lesions, American Journal of Clinical Pathology, Volume 116, Issue 1, July 2001, Pages 17–24, https://doi.org/10.1309/5WFQ-N12E-DT05-UX1T
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Abstract
We describe a novel 4-color polymerase chain reaction (PCR) assay combined with GeneScan analysis to assess for T-cell receptor gamma chain gene (TCRgamma) rearrangements and evaluate its usefulness in 86 lymphoproliferative lesions. In this assay, each variable region (Vgamma) family primer is 5' end-labeled with a different fluorescent dye, allowing determination of the Vgamma family involved in each TCRgamma rearrangement. PCR products were analyzed by capillary electrophoresis. We detected clonal TCRgamma rearrangements in 60 (98%) of 61 T-cell lymphomas, 2 (15%) of 13 B-cell lymphomas, and 3 (25%) of 12 reactive lesions. These results compared favorably with conventional PCR methods using denaturing gradient gel electrophoresis, which revealed clonal TCRgamma rearrangements in 37 (90%) of 41 T-cell lymphomas, 1 (25%) of 4 B-cell lymphomas, and 2 (25%) of 8 reactive lesions. This 4-color PCR assay is at least equivalent to conventional PCR methods and is convenient, allows accurate size determination of TCRgamma rearrangements, and identifies the specific Vgamma family involved, providing more specific information about TCRgamma rearrangement.
Author notes
Dr Vega is a postdoctoral fellow at the University of Texas M.D. Anderson Cancer Center. He is supported by a grant from Fundacion Pedro Barrie de la Maza, Galicia, Spain.