Smitalova Dagmar, Dvorakova Dana, Racil Zdenek, Romzova Marianna
Department of Molecular Medicine, Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
Department of Internal Medicine, Hematology and Oncology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
Pract Lab Med. 2021 Mar 9;25:e00210. doi: 10.1016/j.plabm.2021.e00210. eCollection 2021 May.
molecular detection using quantitative PCR (qPCR) methods is the golden standard of chronic myeloid leukemia (CML) monitoring. However, due to variable sensitivity of qPCR assays across laboratories, alternative methods are tested. Digital PCR (dPCR) has been suggested as a robust and reproducible option. Here we present a comparison of droplet dPCR with routinely used reverse-transcription qPCR (RT-qPCR) and automated GeneXpert systems. Detection limit of dPCR was above 3 BCR copies, although due to background amplification the resulting sensitivity was 0.01% (MR4.0). Nevertheless, in comparison with GeneXpert, dPCR categorized more than 50% of the patients into different MR groups, showing a potential for improved detection.
使用定量聚合酶链反应（qPCR）方法进行分子检测是慢性髓性白血病（CML）监测的金标准。然而，由于各实验室qPCR检测的灵敏度存在差异，人们开始测试替代方法。数字PCR（dPCR）被认为是一种可靠且可重复的选择。在此，我们对液滴dPCR与常规使用的逆转录qPCR（RT-qPCR）及自动化GeneXpert系统进行了比较。dPCR的检测限高于3个BCR拷贝，尽管由于背景扩增，其最终灵敏度为0.01%（MR4.0）。然而，与GeneXpert相比，dPCR将超过50%的患者归入不同的MR组，显示出其在提高检测方面的潜力。
