一种内部 PCR 检测与 BIOMED-2 检测在 T 细胞克隆性评估中的平行比较，为实现高效和具有成本效益的策略提供了依据。

A parallel comparison of T-cell clonality assessment between an in-house PCR assay and the BIOMED-2 assay leading to an efficient and cost-effective strategy.

机构信息

Department of Pathology, Chi-Mei Medical Center, Tainan, Taiwan.

出版信息

J Clin Pathol. 2011 Jun;64(6):536-42. doi: 10.1136/jcp.2010.086637. Epub 2011 Apr 13.

DOI:10.1136/jcp.2010.086637

PMID:21490377

Abstract

AIMS

Diagnosis of T-cell lymphoproliferation is sometimes challenging, and in certain instances pathologists rely heavily on the clonality assessment results of T-cell receptor (TCR) gene rearrangement (TCR-GR). Many investigators have designed various in-house primer sets for PCR-based study targeting different loci of TCR genes. In recent years, the commercial BIOMED-2 protocols have become available. The in-house primers are very cheap while the BIOMED-2 primers are expensive. This parallel study aimed to compare the sensitivity of the in-house TCRG primers (two reactions) and the BIOMED-2 TCR primers (six reactions) in an attempt to develop a sensitive and cost-effective strategy for TCR-GR assessment.

METHODS

PCR-based analysis was performed on 69 samples of T-lineage neoplasms including 60 formalin-fixed paraffin-embedded (FFPE) tissues, 5 samples from peripheral blood (PB) and 4 samples from bone marrow (BM) aspirate.

RESULTS

Forty-seven (78%) FFPE and all PB or BM aspirate samples yielded control DNA products suitable for clonality assessment including 4 precursor and 50 mature T-cell neoplasms. The detection rates of clonal TCR-GR were 63% (34/54) by the two in-house TCRG primers, 85% (46/54) by all six BIOMED-2 reactions, 91% (49/54) by combining the in-house and BIOMED-2 TCRG reactions and 94% (51/54) by combining the in-house and all BIOMED-2 reactions. By using the in-house and BIOMED-2 TCRG reactions with a total of four tubes, clonal TCR-GR was detected in 91% of the cases. The reagent cost for this combination was one-third of that for the six BIOMED-2 reactions and the detection rate was also higher than the latter alone (91% vs 85%).

CONCLUSIONS

As the in-house primers were custom made and are much cheaper than the commercial kits, the authors concluded that this four-tube strategy was cost-effective and efficient for TCR-GR clonality assessment.

摘要

目的

T 细胞淋巴增生的诊断有时具有挑战性，在某些情况下，病理学家严重依赖 T 细胞受体（TCR）基因重排（TCR-GR）的克隆性评估结果。许多研究人员已经设计了用于基于 PCR 的研究的各种内部引物组，针对 TCR 基因的不同基因座。近年来，商业 BIOMED-2 方案已经可用。内部引物非常便宜，而 BIOMED-2 引物则很昂贵。这项平行研究旨在比较内部 TCRG 引物（两个反应）和 BIOMED-2 TCR 引物（六个反应）的敏感性，试图开发一种敏感且具有成本效益的 TCR-GR 评估策略。

方法

对包括 60 例福尔马林固定石蜡包埋（FFPE）组织、5 例外周血（PB）和 4 例骨髓抽吸物在内的 69 例 T 系肿瘤样本进行基于 PCR 的分析。

结果

47 例（78%）FFPE 和所有 PB 或 BM 抽吸物样本均产生适合克隆性评估的对照 DNA 产物，包括 4 例前体细胞和 50 例成熟 T 细胞肿瘤。通过两个内部 TCRG 引物，克隆 TCR-GR 的检出率为 63%（34/54），通过所有六个 BIOMED-2 反应，检出率为 85%（46/54），通过组合内部和 BIOMED-2 TCRG 反应，检出率为 91%（49/54），通过组合内部和所有 BIOMED-2 反应，检出率为 94%（51/54）。使用内部和 BIOMED-2 TCRG 反应总共四个管，91%的病例中检测到克隆 TCR-GR。该组合的试剂成本仅为六个 BIOMED-2 反应的三分之一，检测率也高于后者（91%比 85%）。