Laboratory for Applied Science and Technology in Health, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz), Prof Algacyr Munhoz Mader 3775 Street, Curitiba, Paraná, ZIP 81350-010, Brazil.
Erasto Gaertner Hospital, Dr. Ovande do Amaral 201 Street, Curitiba, Paraná, ZIP 81520-060, Brazil.
Anal Biochem. 2022 Mar 15;641:114541. doi: 10.1016/j.ab.2021.114541. Epub 2021 Dec 29.
The detection of BCR-ABL1 mRNA transcripts is essential to molecular chronic myeloid leukemia (CML) diagnosis. In most cases, the RT-qPCR technique is performed as the gold standard diagnosis tool for clinical cases. However, this method requires expensive reagents and equipment, such as a real-time thermal cycler, probes and master mix. Consequently, the development and validation of simple and low-cost methods are essential for a rapid CML diagnosis in less specialized and equipped centers. In this study, we develop and demonstrate an accessible, rapid, and low-cost method using RT-LAMP for BCR-ABL1 detection in both cell lines and CML clinical samples, using fluorescent and colorimetric assays. Both methods demonstrated diagnostic specificity of 100% and while diagnostic sensitivity reaches more than 90% in samples with RT-qPCR cycle threshold above 31. The obtained data indicates that the proposed method here described is robust, specific and rapid approach for CML diagnosis with outstanding performance, especially for CML diagnostic procedure where present high BCR-ABL1 expression.
检测 BCR-ABL1 mRNA 转录本对于分子慢性髓性白血病 (CML) 的诊断至关重要。在大多数情况下，RT-qPCR 技术是作为临床病例的金标准诊断工具进行的。然而，这种方法需要昂贵的试剂和设备，如实时热循环仪、探针和主混合物。因此，开发和验证简单且低成本的方法对于在较少专业化和配备的中心进行快速 CML 诊断至关重要。在这项研究中，我们开发并展示了一种使用 RT-LAMP 的简单、快速且低成本的方法，用于通过荧光和比色测定法检测细胞系和 CML 临床样本中的 BCR-ABL1。两种方法的诊断特异性均为 100%，而在 RT-qPCR 循环阈值高于 31 的样本中，诊断灵敏度达到 90%以上。所得数据表明，这里描述的方法是一种稳健、特异且快速的 CML 诊断方法，具有出色的性能，特别是对于 BCR-ABL1 高表达的 CML 诊断程序。
