Department of Pathology, Albert Einstein College of Medicine/Montefiore Medical Center, 111 E. 210th Street, Bronx, NY 10467, USA.
J Mol Diagn. 2010 Mar;12(2):226-37. doi: 10.2353/jmoldx.2010.090042.
Detecting clonal T-cell receptor (TCR)-gamma gene rearrangements (GRs) is an important adjunct test for diagnosing T-cell lymphoma. We compared a recently described assay (BIOMED-2 protocol), which targets multiple variable (V) gene segments in two polymerase chain reaction (PCR) reactions (multi-V), with a frequently referenced assay that targets a single V gene segment in four separate PCR reactions (mono-V). A total of 144 consecutive clinical DNA samples were prospectively tested for T-cell clonality by PCR using laboratory-developed mono-V and commercial multi-V primer sets for TCR-gamma GR. The combination of TCR-beta, mono-V TCR-gamma and multi-V TCR-gamma detected more clonal cases (68/144, 47%) than any individual PCR assay. We detected clonal TCR-beta GR in 47/68 (69%) cases. Using either mono-V or multi-V TCR-gamma primers, the sensitivities for detecting clonality were 52/68 (76%) or 51/68 (75%). Using both mono-V and multi-V TCR-gamma primers improved the sensitivity for detecting clonality, 60/68 (88%). Combining either mono-V or multi-V TCR-gamma primers with TCR-beta primers also improved the sensitivity, 64/68 (94%). Significantly, TCR-gamma V11 GRs could only be detected using the mono-V-PCR primers. We conclude that using more than one T-cell PCR assay can enhance the overall sensitivity for detecting T-cell clonality.
检测克隆性 T 细胞受体 (TCR)-γ 基因重排 (GR) 是诊断 T 细胞淋巴瘤的重要辅助检测手段。我们比较了一种新描述的检测方法（BIOMED-2 方案），该方法在两个聚合酶链反应 (PCR) 反应中靶向多个可变 (V) 基因片段（多 V），与一种经常参考的检测方法，该方法在四个单独的 PCR 反应中靶向单个 V 基因片段（单 V）。共前瞻性地检测了 144 例连续的临床 DNA 样本，采用实验室开发的单 V 和商业多 V TCR-γ GR 引物对通过 PCR 检测 T 细胞克隆性。TCR-β、单 V TCR-γ 和多 V TCR-γ 的组合检测到更多的克隆病例（68/144，47%）比任何单个 PCR 检测都多。我们在 47/68（69%）例中检测到克隆性 TCR-β GR。使用单 V 或多 V TCR-γ 引物，检测克隆性的敏感性分别为 52/68（76%）或 51/68（75%）。同时使用单 V 和多 V TCR-γ 引物可提高检测克隆性的敏感性，60/68（88%）。将单 V 或多 V TCR-γ 引物与 TCR-β 引物结合使用也可提高敏感性，64/68（94%）。重要的是，仅使用单 V-PCR 引物才能检测到 TCR-γ V11 GRs。我们得出结论，使用多个 T 细胞 PCR 检测可提高检测 T 细胞克隆性的总体敏感性。
