Evaluation of the cepheid GeneXpert BCR-ABL assay
Evaluation of the cepheid GeneXpert BCR-ABL assay
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DOI：
10.2353/jmoldx.2007.060112
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发表时间：
2007-04-01
影响因子：
4.1
通讯作者：
Gocke, Christopher D.
中科院分区：
文献类型：
--
作者：
Jobbagy, Zsolt;van Atta, Reuel;Gocke, Christopher D.
慢性粒细胞白血病患者存在t(9；22)易位，相当于bcr和abl基因在DNA水平上的融合。翻译的融合产物是一种癌基因蛋白，具有升高的ABL酪氨酸激酶活性，导致细胞转化。到目前为止，逆转录聚合酶链式反应被认为是检测bcr-abl基因融合低拷贝数最敏感的方法。最近，造父变种推出了基于GeneXpert的检测方法，用于从血液样本中鉴定细胞中的bcr-abl基因融合。该系统包括一个独立的仪器，它结合了基于墨盒的微流控样品制备、基于逆转录聚合酶链式反应的荧光信号检测以及BCR-ABL和ABL Ct(阈值周期)的测定。BCR-ABL Ct和ABL Ct(Delta Ct)之间的差异有望代表这两个群体的mRNAs的比率，并最终代表存在的肿瘤细胞的百分比。我们测试了该BCR-ABL融合检测系统是否可以作为监测慢性粒细胞白血病微小残留病患者的临床诊断工具。我们报告了这种自动化的BCR-ABL融合检测系统的性能特征，包括检测极限、特异性、灵敏度和精密度，与基于TaqMan逆转录酶-聚合酶链式反应的手动检测系统相似。
Patients with chronic myeloid leukemia harbor the chromosomal translocation t(9;22), which corresponds to fusion of the BCR and ABL genes at the DNA level. The translated fusion product is an oncogenic protein with increased ABL tyrosine kinase activity causing cell transformation. To date, reverse transcriptase-polymerase chain reaction is considered the most sensitive method available for detecting low copy numbers of the BCR-ABL gene fusion. Recently, Cepheid introduced its GeneXpert-based assay for the identification of the BCR-ABL gene fusion in cells from blood samples. This system comprises a walkaway self-contained instrument that combines cartridge-based microfluidic sample preparation with reverse transcriptase-polymerase chain reaction-based fluorescent signal detection and BCR-ABL and ABL Ct (threshold cycle) determination. The difference between the BCR-ABL Ct and ABL Ct (Delta Ct) is expected to represent the ratio of the two populations of mRNAs and ultimately the percentage of neoplastic cells present. We tested whether this BCR-ABL fusion detection system could be used as a clinical diagnostic tool for monitoring patients with minimal residual disease of chronic myelogenous leukemia. We report similar performance characteristics, including limit of detection, specificity, sensitivity, and precision, of this automated BCR-ABL fusion detection system to those of a manual TaqMan reverse transcriptase-polymerase chain reaction-based test.