Assessment of a new RNA stabilizing reagent (Tempus Blood RNA) for minimal residual disease in onco-hematology using the EAC protocol
Assessment of a new RNA stabilizing reagent (Tempus Blood RNA) for minimal residual disease in onco-hematology using the EAC protocol
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DOI：
10.1016/j.leukres.2005.08.027
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发表时间：
2006-05-01
影响因子：
2.7
通讯作者：
Picard, C
中科院分区：
文献类型：
--
作者：
Prezeau, N;Silvy, M;Picard, C
实时定量聚合酶链式反应(RQ-PCR)的标准化工作，如欧洲抗癌(EAC)协议，实际上是必不可少的。然而，分析前阶段的所有阶段(血液采集。保守性和细胞分离程序)，影响许多基因的体外表达，但不受控制。开发了在采集血液时稳定RNA的各种试剂盒：PAX基因血液试剂盒(Qigen)和新的Tempus血液RNA试剂盒(应用生物系统)。PAX基因血液已经被证实可以监测肿瘤血液学中的微小残留病(MRD)。为了对Tempus血液RNA试剂盒进行评价，将其与未稳定的EDTA血液/Ficoll/TRIzol方案和PAX基因血液试剂盒进行了比较。对不同方法提取的RNA的数量、质量和控制基因(GUS和ABL)以及融合基因转录本BCR-ABL和TEL-AML1的检测进行了RQ-PCR分析。我们的研究表明，新的Tempus血液RNA试剂盒使用EAC方案，可以以与其他两种方案相同的灵敏度检测融合基因转录物。总之，我们的数据表明，这种技术可能用于骨髓增生性疾病和急性白血病的MRD随访。因此，应该对白血病患者进行进一步的多中心研究，以验证这项技术在这些应用中的有效性。(C)2005爱思唯尔有限公司。保留所有权利。
Standardization efforts of real time quantitative PCR (RQ-PCR), such as the Europe Against Cancer (EAC) protocol, are actually essential. However, all the stages of the preanalytical phase (blood collection. conservation and procedures of cellular separation) which influence the ex vivo expression of many genes are not controlled. Various kits for stabilizing the RNA at the time of blood collection were developed: PAXgene Blood kit (Qiagen) and the new Tempus Blood RNA kit (Applied Biosystems). PAXgene Blood was already validated to monitor minimal residual disease (MRD) in onco-hematologic pathologies. In order to evaluate the Tempus Blood RNA kit, it was compared to unstabilized EDTA blood/Ficoll/TRIzol protocol and PAXgene Blood kit. The RNAs extracted by the different methods were assessed for quantity, quality and detection of control genes (GUS and ABL) and fusion gene transcripts BCR-ABL and TEL-AML1 by RQ-PCR.Our study shows that using the EAC protocol, the new Tempus Blood RNA kit allows the detection of fusion gene transcript with the same sensitivity as the two other protocols. Altogether, our data suggest the possible use of such a technology for MRD follow-up in myeloproliferative diseases and acute leukemias. So, further multicentric studies on leukemic patients should be performed to validate this technique in these applications. (C) 2005 Elsevier Ltd. All rights reserved.