Chabi Maede, Vu Binh, Brosamer Kristen, Song Sophia, Maranholkar Vijay, Zeng Zihua, Zu Youli, Kanagal-Shamanna Rashmi, Conrad Jacinta C, Willson Richard C, Kourentzi Katerina
Department of Biomedical Engineering, University of Houston Houston Texas 77204 USA
Department of Chemical and Biomolecular Engineering, University of Houston Houston Texas 77204 USA
Sens Diagn. 2025 Mar 21;4(5):416-424. doi: 10.1039/d4sd00357h. eCollection 2025 May 15.
Due to the slow progression of most cancers, speed of diagnosis is not of primary concern. However, the diagnosis of acute promyelocytic leukemia (APL) is unusually urgent because its hemorrhagic complications can result in death within a few days. APL is highly treatable, but the turnaround time for standard molecular testing often exceeds the window for life-saving treatment, even in advanced medical centers. The hallmark of APL is the fusion of the PML and RARα genes (t(15;17)) resulting in the expression of a growth-promoting PML-RARα fusion protein. Toward timely screening for APL, we have developed a sensitive europium-based lateral flow immunoassay for direct detection of nuclear PML-RARα fusion oncoprotein. We demonstrated a limit of detection of 11% fusion protein positive NB4 cells spiked into healthy peripheral blood mononuclear cells and an integrated filter-based sample preparation workflow showcasing its potential for clinically actionable utility in prompt APL screening. With further validation with clinical human samples this lateral flow immunoassay has the potential to enable fusion-protein based cancer diagnostics at true point-of-care.
由于大多数癌症进展缓慢，诊断速度并非首要关注点。然而，急性早幼粒细胞白血病（APL）的诊断异常紧急，因为其出血性并发症可在数天内导致死亡。APL具有高度可治性，但即使在先进的医疗中心，标准分子检测的周转时间往往超过挽救生命治疗的时间窗。APL的标志是PML和RARα基因融合（t(15;17)），导致促生长的PML-RARα融合蛋白表达。为了及时筛查APL，我们开发了一种基于铕的灵敏侧向流动免疫分析法，用于直接检测核PML-RARα融合癌蛋白。我们证明，将11%融合蛋白阳性的NB4细胞加入健康外周血单核细胞中的检测限，并展示了基于滤膜的集成样本制备工作流程，突显了其在快速APL筛查中临床可操作实用性的潜力。通过临床人体样本的进一步验证，这种侧向流动免疫分析法有可能在真正的即时检测点实现基于融合蛋白的癌症诊断。
