Department of Laboratory Medicine, Med+X Center for Manufacturing, West China Precision Medicine Industrial Technology Institute, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, China; Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Chongqing, 402160, China; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Anal Chim Acta. 2023 Oct 2;1276:341623. doi: 10.1016/j.aca.2023.341623. Epub 2023 Jul 13.
Acute promyelocytic leukemia (APL) is an acute myeloid leukemia (AML) with a specific fusion gene target, PML/RARα fusion gene (PML/RARα), which is formed by the translocation of chromosomes 15 and 17. Detection of PML/RARα is the most reliable parameter for the diagnosis, treatment adjustment, efficacy evaluation, prognosis analysis and relapse prediction of APL. In this study, a novel biosensor was constructed for rapid enzyme-free detection of PML/RARα using DNAzyme and carbon dots/cobalt oxhydroxide nanosheet complexs (CDs/CoOOH). In the detection system, the separated DNAzymes could specifically recognize and bind together by the PML/RARα to form a complete DNAzyme for shearing hairpin probe (HP), then generated trigger, which was the first signal amplification. Then, trigger could hybridize with the capture probe (CP) anchored to streptavidin (SA) modified microplate as well as fluorescence quenching signal probe (SP@CDs/CoOOH). Finally, ascorbic acid (AA) was added to decompose CoOOH and the fluorescence of CDs was released, which was the second signal amplification. Through the dual signal amplification of DNAzyme and CDs/CoOOH, PML/RARα could be detected quickly and sensitively, which overcame the limitation of protein enzyme in traditional fluorescence methods, showing potential clinical application value in the diagnosis and treatment of leukemia.
急性早幼粒细胞白血病（APL）是一种具有特定融合基因靶点的急性髓细胞白血病（AML），即 PML/RARα 融合基因（PML/RARα），它是由染色体 15 和 17 的易位形成的。检测 PML/RARα 是 APL 诊断、治疗调整、疗效评估、预后分析和复发预测最可靠的参数。在这项研究中，构建了一种新型的生物传感器，用于使用 DNAzyme 和碳点/钴氢氧化物纳米片复合物（CDs/CoOOH）快速酶免检测 PML/RARα。在检测系统中，分离的 DNAzyme 可以通过 PML/RARα 特异性识别和结合在一起，形成完整的 DNAzyme 来剪切发夹探针（HP），从而产生触发物，这是第一个信号放大。然后，触发物可以与固定在链霉亲和素（SA）修饰的微孔板上的捕获探针（CP）以及荧光猝灭信号探针（SP@CDs/CoOOH）杂交。最后，加入抗坏血酸（AA）分解 CoOOH，释放出 CDs 的荧光，这是第二个信号放大。通过 DNAzyme 和 CDs/CoOOH 的双重信号放大，可以快速、灵敏地检测 PML/RARα，克服了传统荧光方法中蛋白质酶的局限性，在白血病的诊断和治疗中具有潜在的临床应用价值。
