College of Chemistry, Guangdong University of Petrochemical Technology, Guandu Road, Maoming, Guangdong, 525000, People's Republic of China.
School of Life and Environmental Sciences, School of Intellectual Property, Guilin University of Electronic Technology, Guilin, Guangxi, 541004, People's Republic of China.
Mikrochim Acta. 2024 Aug 20;191(9):547. doi: 10.1007/s00604-024-06628-2.
Atherosclerosis cardiovascular disease (ASCVD) has become one of the leading death causes in humans. Low-density lipoprotein (LDL) is an important biomarker for assessing ASCVD risk level. Thus, monitoring LDL levels can be an important means for early diagnosis of ASCVD. Herein, a novel electrochemical aptasensor for determination LDL was designed based on nitrogen-doped reduced graphene oxide-hemin-manganese oxide nanoparticles (NrGO-H-MnO NPs) integrated with clustered regularly interspaced short palindromic repeats and associated proteins (CRISPR/Cas12a) system. NrGO-H-MnO NPs not only have a large surface area and remarkable enhanced electrical conductivity but also the interconversion of different valence states of iron in hemin can provide an electrical signal. Nonspecific single-stranded DNA (ssDNA) was bound to NrGO-H-MnO NPs to form a signaling probe and was immobilized on the electrode surface. The CRISPR/Cas12a system has excellent trans-cleavage activity, which can be used to cleave ssDNA, thus detaching the NrGO-H-MnO NPs from the sensing interface and attenuating the electrical signal. Significant signal change triggered by the target was ultimately obtained, thus achieving sensitive detection of the LDL in range from 0.005 to 1000.0 nM with the detection limit of 0.005 nM. The proposed sensor exhibited good stability, selectivity, and stability and achieved reliable detection of LDL in serum samples, demonstrating its promising application prospects for the diagnostic application of LDL.
动脉粥样硬化性心血管疾病（ASCVD）已成为人类主要死亡原因之一。低密度脂蛋白（LDL）是评估 ASCVD 风险水平的重要生物标志物。因此，监测 LDL 水平可以作为 ASCVD 早期诊断的重要手段。在此，设计了一种基于氮掺杂还原氧化石墨烯-血红素-氧化锰纳米粒子（NrGO-H-MnO NPs）与成簇规律间隔短回文重复序列和相关蛋白（CRISPR/Cas12a）系统的新型用于测定 LDL 的电化学适体传感器。NrGO-H-MnO NPs 不仅具有较大的表面积和显著增强的导电性，而且血红素中铁的不同价态的相互转换可以提供电信号。非特异性单链 DNA（ssDNA）与 NrGO-H-MnO NPs 结合形成信号探针，并固定在电极表面。CRISPR/Cas12a 系统具有出色的转录切割活性，可用于切割 ssDNA，从而将 NrGO-H-MnO NPs 从传感界面上脱离并减弱电信号。最终会产生由靶标触发的显著信号变化，从而实现了在 0.005 至 1000.0 nM 范围内对 LDL 的灵敏检测，检测限为 0.005 nM。所提出的传感器表现出良好的稳定性、选择性和重现性，并且可以可靠地检测血清样品中的 LDL，表明其在 LDL 诊断应用方面具有广阔的应用前景。
