文章：

建立具有国际报告标准BCR-ABL1定量测量所需分析性能的标准化多重分析方法

Establishment of a standardized multiplex assay with the analytical performance required for quantitative measurement of BCR–ABL1 on the international reporting scale

原文发布日期：2011-03-25

DOI: 10.1038/bcj.2011.10

类型: Original Article

开放获取: 是

英文摘要：

Accurate and standardized methods for the quantitative measurement of BCR–ABL1 are a prerequisite for monitoring of treatment response in t(9;22)-positive leukemia. Here, we describe a novel multiplex assay system based on the proven TaqMan and Armored RNA technologies and optimized for sensitive detection of three BCR–ABL1 fusion transcripts and ABL1 in a single reaction. Analytical experiments confirmed the absence of significant competition between the simultaneous amplification reactions and established the sensitivity, linearity and precision of the assay. Comparative studies with 115 clinical specimens resulted in high qualitative and quantitative agreement with independent singleplex laboratory-developed tests routinely used in clinical testing. Direct comparison with a reference laboratory calibrated to the international scale (IS) demonstrated minimal analytical bias between methods and an overall accuracy and precision within the performance range required for quantitative measurement of BCR–ABL1 on the IS. We conclude that detection of e1a2, b2a2, b3a2 and ABL1 can be achieved in a multiplex assay format compatible with IS reporting. Further clinical validation of the assay could improve the operational efficiency of clinical laboratories, increase their adherence to current recommendations for b2a2/b3a2 reporting on the IS and provide for the first time an opportunity to standardize e1a2-monitoring results.

摘要翻译： 

准确且标准化的BCR-ABL1定量检测方法是监测t(9;22)阳性白血病治疗反应的前提。本文基于成熟的TaqMan技术和Armored RNA技术，开发了一种新型多重检测系统，可在单次反应中同步检测三种BCR-ABL1融合转录本和ABL1，并实现灵敏检测。分析实验证实同步扩增反应间无显著竞争，并验证了该检测方法的灵敏度、线性及精密度。对115例临床标本的对比研究显示，与临床检测常规使用的独立单重实验室自建检测方法在定性和定量方面高度一致。与国际标度（IS）校准的参考实验室直接比较表明，方法间分析偏差极小，整体准确度和精密度均符合IS体系下BCR-ABL1定量检测的性能要求。我们得出结论：e1a2、b2a2、b3a2和ABL1的检测可通过兼容IS报告的多重检测形式实现。该检测方法的进一步临床验证可提升临床实验室运营效率，促进b2a2/b3a2的IS报告更符合现行指南要求，并首次为e1a2监测结果的标准化提供可能。

原文链接：

Establishment of a standardized multiplex assay with the analytical performance required for quantitative measurement of BCR–ABL1 on the international reporting scale