Lamberson C, Hutchison R E, Shrimpton A E
Department of Pathology, State University of New York Upstate Medical University, 750 E Adams St., Syracuse, NY 13210, USA.
Mol Diagn. 2001 Jun;6(2):117-24. doi: 10.1054/modi.2001.25321.
The T-cell receptor gamma chain (TCR-gamma) gene has 11 functional variable (V) exons that can be organized into four subfamilies and four functional joining (J) exons that can be divided into two subfamilies.
Three multiplex PCR reactions amplifying the TCR-gamma gene were developed. Primer combinations for multiplex PCR were chosen so that the V-region subfamily and J-region subfamily involved in a clonal band could be identified. Control primers from the protease inhibitor (PI) gene were also included in each reaction to verify the presence of amplifiable DNA. Fifty-six archived samples that had been tested by Southern blot for clonal rearrangement of the TCR-beta gene were analyzed with the TCR-gamma PCR protocol. Twenty-one of 56 samples were TCR-beta positive by Southern blot and thus expected to be positive with TCR-gamma PCR. Thirty-five of 56 samples were TCR-beta negative by Southern blot. Of these, 14 samples showed clonal rearrangement of the immunoglobulin heavy chain gene. The TCR-gamma PCR protocol showed a diagnostic sensitivity for detecting T-lineage clonality of 90%, with a diagnostic specificity for detecting T-cell lineage of only 74%.
The PCR protocol described here performed well in comparison with a TCR-beta Southern protocol.
T细胞受体γ链（TCR-γ）基因有11个功能性可变（V）外显子，可分为四个亚家族，还有四个功能性连接（J）外显子，可分为两个亚家族。
开发了三种扩增TCR-γ基因的多重PCR反应。选择多重PCR的引物组合，以便能够识别克隆条带中涉及的V区亚家族和J区亚家族。每个反应中还包含来自蛋白酶抑制剂（PI）基因的对照引物，以验证可扩增DNA的存在。采用TCR-γ PCR方案分析了56份已通过Southern印迹法检测TCR-β基因克隆重排的存档样本。56份样本中有21份通过Southern印迹法检测为TCR-β阳性，因此预期TCR-γ PCR检测也为阳性。56份样本中有35份通过Southern印迹法检测为TCR-β阴性。其中，14份样本显示免疫球蛋白重链基因的克隆重排。TCR-γ PCR方案检测T系克隆性的诊断敏感性为90%，而检测T细胞系的诊断特异性仅为74%。
与TCR-β Southern方案相比，本文所述的PCR方案表现良好。
