Wang Zhilong, Wei Luyu, Ruan Shilong, Chen Yiping
College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China.
Daye Public Inspection and Test Center, Daye, Hubei 435100, People's Republic of China.
J Agric Food Chem. 2023 Mar 15;71(10):4417-4425. doi: 10.1021/acs.jafc.3c00341. Epub 2023 Feb 28.
Herein, we report a CRISPR/Cas12a-assisted chemiluminescence sensor for aflatoxin B (AFB) detection based on functional nucleic-acid-mediated target recognition and in-pipet rolling circle amplification-mediated signal amplification. In this sensor, we performed rolling circle amplification on the inside of the pipet to enrich horseradish peroxidase (pipet-poly-HRP). When AFB is present, it interacts with functional nucleic acids and results in the release of the activator. The activator is designed to activate the CRISPR/Cas12a system, which cleaves the pipet-poly-HRP to liberate HRP. The freed HRP can then be measured by chemiluminescence to quantify AFB. This CRISPR/Cas12a-assisted chemiluminescence sensor enables facile, highly sensitive, and specific detection of AFB, with a linear range from 50 pg/mL to 100 ng/mL and a detection limit of 5.2 pg/mL. Furthermore, it exhibits satisfactory recovery and has successfully challenged AFB detection in cereal samples. The proposed sensor offers a novel rapid screening approach that holds great promise for food security monitoring.
在此，我们报道了一种基于功能核酸介导的靶标识别和移液管内滚环扩增介导的信号放大的CRISPR/Cas12a辅助化学发光传感器，用于黄曲霉毒素B（AFB）检测。在该传感器中，我们在移液管内部进行滚环扩增以富集辣根过氧化物酶（移液管-聚-HRP）。当存在AFB时，它与功能核酸相互作用并导致激活剂释放。激活剂旨在激活CRISPR/Cas12a系统，该系统切割移液管-聚-HRP以释放HRP。然后可以通过化学发光测量释放的HRP来定量AFB。这种CRISPR/Cas12a辅助化学发光传感器能够简便、高灵敏且特异检测AFB，线性范围为50 pg/mL至100 ng/mL，检测限为5.2 pg/mL。此外，它具有令人满意的回收率，并已成功用于谷物样品中AFB的检测。所提出的传感器提供了一种新颖的快速筛查方法，对食品安全监测具有巨大潜力。
