Neuroelectronics, Munich Institute of Biomedical Engineering, Department of Electrical Engineering, School of Computation, Information and Technology, Technical University of Munich, 85748 Garching, Germany.
Institute of Biological Information Processing, Bioelectronics (IBI-3), Forschungszentrum Jülich, 52425 Jülich, Germany.
ACS Sens. 2024 Nov 22;9(11):6197-6206. doi: 10.1021/acssensors.4c02060. Epub 2024 Oct 22.
The rapid and reliable detection and quantification of nucleic acids is crucial for various applications, including infectious disease and cancer diagnostics. While conventional methods, such as the quantitative polymerase chain reaction are widely used, they are limited to the laboratory environment due to their complexity and the requirement for sophisticated equipment. In this study, we present a novel amplification-free digital sensing strategy by combining the collateral cleavage activity of the Cas12a enzyme with single-impact electrochemistry. In doing so, we modified silver nanoparticles using a straightforward temperature-assisted cofunctionalization process to subsequently detect the collision events of particles released by the activated Cas12a as distinct current spikes on a microelectrode array. The functionalization resulted in stable DNA-AgNP conjugates, making them suitable for numerous biosensor applications. Thus, our study demonstrates the potential of clustered regularly interspaced short palindromic repeats-based diagnostics combined with impact-based digital sensing for a rapid and amplification-free quantification of nucleic acids.
核酸的快速可靠检测和定量对于各种应用至关重要，包括传染病和癌症诊断。虽然传统方法，如定量聚合酶链反应，由于其复杂性和对复杂设备的要求，广泛应用于实验室环境中。在这项研究中，我们提出了一种新颖的无扩增数字传感策略，将 Cas12a 酶的旁切活性与单冲击电化学结合起来。为此，我们通过简单的温度辅助共功能化过程对银纳米粒子进行了修饰，随后在微电极阵列上检测到由激活的 Cas12a 释放的粒子的碰撞事件，表现为明显的电流尖峰。该功能化导致了稳定的 DNA-AgNP 缀合物，使其适用于许多生物传感器应用。因此，我们的研究表明，基于聚类规则间隔短回文重复序列的诊断与基于冲击的数字传感相结合，可用于快速无扩增定量核酸。
