人乳头瘤病毒 (HPV) 的持续感染会显着促进宫颈癌的发生。因此，迫切需要开发快速、准确的HPV检测方法。在此，我们提出了一种基于 CRISPR/Cas12a 的超灵敏电化学发光 (ECL) 成像技术，用于检测 HPV-18 DNA。 ECL DNA 传感器阵列是通过应用黑洞猝灭剂 (BHQ) 和聚合物点 (Pdots) 共标记构建的将发夹 DNA (hpDNA) 固定到镀金氧化铟锡载玻片 (Au-ITO) 上。 ECL 成像方法包括将目标 HPV-18 与 crRNA 和 Cas12a 的混合物一起孵育以激活 Cas12a，然后将活性 Cas12a 与 ECL 传感器一起孵育。这种相互作用通过消化传感器表面的 hpDNA 导致 BHQ 从 Pdot 上无差别裂解，从而恢复 Pdot 的 ECL 信号。 ECL 亮度读数展示了 ECL 成像技术的卓越性能，线性检测范围为 10 fM-500 pM，检测限 (LOD) 为 5.3 fM。基于 Cas12a 的 ECL 成像方法具有高灵敏度和检测范围广泛，使其在核酸检测应用中极具前景。版权所有 © 2024 Elsevier B.V. 保留所有权利。

Persistent infection with human papillomavirus (HPV) significantly contributes to the development of cervical cancer. Thus, it is urgent to develop rapid and accurate methods for HPV detection. Herein, we present an ultrasensitive CRISPR/Cas12a-based electrochemiluminescent (ECL) imaging technique for the detection of HPV-18 DNA.The ECL DNA sensor array is constructed by applying black hole quencher (BHQ) and polymer dots (Pdots) co-labeled hairpin DNA (hpDNA) onto a gold-coated indium tin oxide slide (Au-ITO). The ECL imaging method involves an incubation process of target HPV-18 with a mixture of crRNA and Cas12a to activate Cas12a, followed by an incubation of the active Cas12a with the ECL sensor. This interaction causes the indiscriminate cleavage of BHQ from Pdots by digesting hpDNA on the sensor surface, leading to the restoration of the ECL signal of Pdots. The ECL brightness readout demonstrates superior performance of the ECL imaging technique, with a linear detection range of 10 fM-500 pM and a limit-of-detection (LOD) of 5.3 fM.The Cas12a-based ECL imaging approach offers high sensitivity and a broad detection range, making it highly promising for nucleic acid detection applications.Copyright © 2024 Elsevier B.V. All rights reserved.