Han Xi-qun, Qi Zong-li, He Li, Lu Di, Zhao Tong
Department of Pathology, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2004 Feb;24(2):188-91.
To explore the value of detecting clonal T cell receptor gamma (TCR-gamma) gene rearrangement with touch-down PCR and single-strand conformational polymorphism analysis (SSCP) in the diagnosis of lymphoid leukemia.
The DNA of peripheral blood leucocytes from lymphoid leukemia patients were extracted for amplification of the TCR-gamma gene rearrangement with the consensus primers and touch-down PCR. The PCR products were analyzed by agarose gel electrophoresis, direct DNA sequencing and SSCP analysis. The positive control cell line DNA was mixed in different proportions with the DNA extracted from reactive lymphoid tissue to test the sensitivity of the touch-down PCR.
Fifteen of 18 T lymphoid leukemia and 2 of the 4 B lymphoid leukemia patients were identified to be positive by agarose electrophoresis. The positive PCR products were further analyzed by SSCP analysis, which showed discrete bands. Direct DNA sequencing confirmed the clones to be TCR-gamma gene rearrangement, and the sensitivity of touch-down PCR was 1%.
Consensus primers for studying TCR-gamma gene rearrangement in combination with touch-down PCR can effectively amplify the clonal TCR-gamma gene rearrangement in T lymphoid leukemia.
探讨采用降落式聚合酶链反应（touch-down PCR）和单链构象多态性分析（SSCP）检测克隆性T细胞受体γ（TCR-γ）基因重排在淋巴样白血病诊断中的价值。
提取淋巴样白血病患者外周血白细胞DNA，用通用引物和降落式PCR扩增TCR-γ基因重排。PCR产物经琼脂糖凝胶电泳、直接DNA测序和SSCP分析。将阳性对照细胞系DNA与反应性淋巴组织提取的DNA按不同比例混合，检测降落式PCR的灵敏度。
18例T淋巴样白血病患者中有15例、4例B淋巴样白血病患者中有2例经琼脂糖电泳鉴定为阳性。对阳性PCR产物进一步进行SSCP分析，显示出离散条带。直接DNA测序证实这些克隆为TCR-γ基因重排，降落式PCR的灵敏度为1%。
用于研究TCR-γ基因重排的通用引物联合降落式PCR可有效扩增T淋巴样白血病中的克隆性TCR-γ基因重排。
