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Detection of clonal T-cell receptor gamma gene rearrangements using fluorescent-based PCR and automated high-resolution capillary electrophoresis.
Mol Diagn. 2001 Sep; 6(3):169-79.MD

Abstract

BACKGROUND

Analysis of T-cell receptor gamma (TCR gamma) gene rearrangements by PCR is a powerful tool for detecting clonal T-cell populations for the diagnosis of lymphoid neoplasms. We report a method for TCR gamma PCR analysis using capillary electrophoresis (CE).

METHODS AND RESULTS

To define the threshold for identification of a predominant monoclonal population within a polyclonal background, we developed a novel objective parameter of the peak height ratio (Rn) of the peak of interest and the average of the two immediate flanking peaks. After evaluation of monoclonal, reactive, and normal T-cell populations, an Rn of 3.0 or greater was determined to be consistent with a monoclonal population, whereas an Rn between 1.9 and 3.0 was considered an intermediate range. This CE method was compared with the standard denaturing gradient gel electrophoresis (DGGE) method using previously evaluated clinical specimens. Eleven of 12 clinical specimens (92%) with a definitive diagnosis of T-cell lymphoma were monoclonal by CE, with 100% concordance with the DGGE method. Of nine specimens morphologically suspicious for T-cell lymphoma, five specimens were positive by CE analysis compared with four specimens by DGGE. In addition, 14 specimens for staging from patients with known T-cell lymphoma were studied using both the CE and DGGE methods, with a concordance of 86%.

CONCLUSION

CE is a powerful and efficient method for analysis of clonality by TCR gamma PCR.

Authors+Show Affiliations

Molecular Pathology Laboratory, Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104-4283, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11571710

Citation

Luo, V, et al. "Detection of Clonal T-cell Receptor Gamma Gene Rearrangements Using Fluorescent-based PCR and Automated High-resolution Capillary Electrophoresis." Molecular Diagnosis : a Journal Devoted to the Understanding of Human Disease Through the Clinical Application of Molecular Biology, vol. 6, no. 3, 2001, pp. 169-79.
Luo V, Lessin SR, Wilson RB, et al. Detection of clonal T-cell receptor gamma gene rearrangements using fluorescent-based PCR and automated high-resolution capillary electrophoresis. Mol Diagn. 2001;6(3):169-79.
Luo, V., Lessin, S. R., Wilson, R. B., Rennert, H., Tozer, C., Benoit, B., & Leonard, D. G. (2001). Detection of clonal T-cell receptor gamma gene rearrangements using fluorescent-based PCR and automated high-resolution capillary electrophoresis. Molecular Diagnosis : a Journal Devoted to the Understanding of Human Disease Through the Clinical Application of Molecular Biology, 6(3), 169-79.
Luo V, et al. Detection of Clonal T-cell Receptor Gamma Gene Rearrangements Using Fluorescent-based PCR and Automated High-resolution Capillary Electrophoresis. Mol Diagn. 2001;6(3):169-79. PubMed PMID: 11571710.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of clonal T-cell receptor gamma gene rearrangements using fluorescent-based PCR and automated high-resolution capillary electrophoresis. AU - Luo,V, AU - Lessin,S R, AU - Wilson,R B, AU - Rennert,H, AU - Tozer,C, AU - Benoit,B, AU - Leonard,D G, PY - 2001/9/26/pubmed PY - 2002/1/5/medline PY - 2001/9/26/entrez SP - 169 EP - 79 JF - Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology JO - Mol Diagn VL - 6 IS - 3 N2 - BACKGROUND: Analysis of T-cell receptor gamma (TCR gamma) gene rearrangements by PCR is a powerful tool for detecting clonal T-cell populations for the diagnosis of lymphoid neoplasms. We report a method for TCR gamma PCR analysis using capillary electrophoresis (CE). METHODS AND RESULTS: To define the threshold for identification of a predominant monoclonal population within a polyclonal background, we developed a novel objective parameter of the peak height ratio (Rn) of the peak of interest and the average of the two immediate flanking peaks. After evaluation of monoclonal, reactive, and normal T-cell populations, an Rn of 3.0 or greater was determined to be consistent with a monoclonal population, whereas an Rn between 1.9 and 3.0 was considered an intermediate range. This CE method was compared with the standard denaturing gradient gel electrophoresis (DGGE) method using previously evaluated clinical specimens. Eleven of 12 clinical specimens (92%) with a definitive diagnosis of T-cell lymphoma were monoclonal by CE, with 100% concordance with the DGGE method. Of nine specimens morphologically suspicious for T-cell lymphoma, five specimens were positive by CE analysis compared with four specimens by DGGE. In addition, 14 specimens for staging from patients with known T-cell lymphoma were studied using both the CE and DGGE methods, with a concordance of 86%. CONCLUSION: CE is a powerful and efficient method for analysis of clonality by TCR gamma PCR. SN - 1084-8592 UR - https://www.unboundmedicine.com/medline/citation/11571710/Detection_of_clonal_T_cell_receptor_gamma_gene_rearrangements_using_fluorescent_based_PCR_and_automated_high_resolution_capillary_electrophoresis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1084-8592(01)67888-3 DB - PRIME DP - Unbound Medicine ER -