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Detection of Clonal T-cell Receptor γ Gene Rearrangements Using Fluorescent-based PCR and Automated High-resolution Capillary Electrophoresis

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Abstract

Background: Analysis of T-cell receptor γ (TCRγ) 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 TCRγ 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 TCRγ PCR.

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Authors and Affiliations

  1. Molecular Pathology Laboratory, Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Vivian Luo MD,MS, Stuart R. Lessin MD, Robert B. Wilson MD, PhD, Hanna Rennert PhD, Cindy Tozer MS, Bernice Benoit BS & Debra G. B. Leonard MD, PhD

  2. Department of Dermatology, University of Pennsylvania, Philadelphia, PA, USA

    Stuart R. Lessin MD

Authors
  1. Vivian Luo MD,MS
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  2. Stuart R. Lessin MD
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  3. Robert B. Wilson MD, PhD
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  4. Hanna Rennert PhD
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  5. Cindy Tozer MS
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  6. Bernice Benoit BS
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  7. Debra G. B. Leonard MD, PhD
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Correspondence to Debra G. B. Leonard MD, PhD.

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Luo, V., Lessin, S.R., Wilson, R.B. et al. Detection of Clonal T-cell Receptor γ Gene Rearrangements Using Fluorescent-based PCR and Automated High-resolution Capillary Electrophoresis. Molecular Diagnosis 6, 169–179 (2001). https://doi.org/10.1007/BF03262050

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  • DOI: https://doi.org/10.1007/BF03262050

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