Department of Clinical Laboratory, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, China.
Department of Central Laboratory, Shanghai Tenth People's Hospital of Tongji University, Shanghai, 200072, China.
Analyst. 2022 Jul 12;147(14):3186-3192. doi: 10.1039/d2an00697a.
Alpha-fetoprotein (AFP) is an important clinical tumor marker of hepatoblastoma, and the concentration of AFP in serum is closely related to the staging of hepatoblastoma. We report a magnetic bead separation platform based on a switching aptamer triggered hybridization chain reaction (SAT-HCR) and the CRISPR-Cas12a sensor for the detection of AFP. AFP aptamer, as an easily regulated nucleic acid strand, is responsible for binding to AFP into nucleic acid detection, while HCR-CRISPR-Cas12a, regulated by functionalized magnetic nanoparticles, is responsible for highly specific nucleic acid signal amplification. Under the optimal conditions, the fluorescence intensity was proportional to the concentration of AFP in the range of 0.5-10 ng mL and the limit of detection was 0.170 ng mL. In addition, we have successfully applied this biosensor to detect AFP in clinical samples from patients with hepatoblastoma, with greater sensitivity relative to ELISA. Our proposed method showed great potential application in clinical diagnosis and pharmaceutical-related fields with the properties of high sensitivity, low cost and high selectivity.
甲胎蛋白（AFP）是肝母细胞瘤的重要临床肿瘤标志物，血清中 AFP 的浓度与肝母细胞瘤的分期密切相关。我们报道了一种基于切换适体触发杂交链式反应（SAT-HCR）和 CRISPR-Cas12a 传感器的磁珠分离平台，用于 AFP 的检测。AFP 适体作为一种易于调控的核酸链，负责与 AFP 结合进行核酸检测，而 HCR-CRISPR-Cas12a 则由功能化的磁性纳米颗粒调控，负责高度特异性的核酸信号放大。在最佳条件下，荧光强度与 AFP 在 0.5-10ng/mL 范围内的浓度呈正比，检测限为 0.170ng/mL。此外，我们已经成功地将这种生物传感器应用于检测肝母细胞瘤患者的临床样本中的 AFP，与 ELISA 相比具有更高的灵敏度。我们提出的方法具有高灵敏度、低成本和高选择性等特点，在临床诊断和药物相关领域具有很大的应用潜力。
