Graduate School of Chemical Sciences and Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo, 060-8628, Japan.
Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo, 060-8628, Japan.
Biosens Bioelectron. 2022 Feb 15;198:113832. doi: 10.1016/j.bios.2021.113832. Epub 2021 Nov 23.
This study aimed to develop an electrochemical system for measuring blood ATP and lactate levels in a single format. The ratio of lactate to ATP levels was previously reported to provide an alternative illness severity score. Although severity evaluation is crucial to treat patients with acute disease admitted to intensive care units, no sensors are currently available to simply and rapidly measure ATP and lactate levels using the same detection method. Therefore, we constructed an integrated sensing system for ATP and lactate using enzymatic reactions and two sets of electrodes integrated into a chip connected to a single potentiostat operated by a microcontroller. The enzymatic system involves adenylate kinase, pyruvate kinase, and pyruvate oxidase for ATP, and lactate oxidase for lactate, both of which produce hydrogen peroxide. Multiplex enzyme-based reactions were designed to minimize the corresponding operations significantly without enzyme immobilization onto the electrodes. The system was robust in the presence of potentially interfering blood components, such as ascorbate, pyruvate, ADP, urate, and potassium ions. The ATP and lactate levels in the blood were successfully measured using the new sensor with good recoveries. The analytical results of blood samples obtained using our sensor were in good agreement with those using conventional methods. Integrating electrode-based analysis and a microcontroller-based system saved further operations, enabling the straightforward measurement of ATP and lactate levels within 5 min. The proposed sensor may serve as a useful tool in the management of serious infectious diseases.
本研究旨在开发一种电化学系统,以单一格式测量血液中的 ATP 和乳酸水平。此前有报道称,乳酸与 ATP 水平的比值可提供替代疾病严重程度评分。尽管严重程度评估对于治疗入住重症监护病房的急性疾病患者至关重要,但目前尚无传感器可使用相同的检测方法简单快速地测量 ATP 和乳酸水平。因此,我们使用酶反应和集成到与单个微控制器操作的电位计相连的芯片中的两组电极构建了用于 ATP 和乳酸的集成传感系统。酶系统涉及用于 ATP 的腺苷酸激酶、丙酮酸激酶和丙酮酸氧化酶,以及用于乳酸的乳酸氧化酶,两者均产生过氧化氢。设计了基于多重酶的反应,在不将酶固定在电极上的情况下,显著减少相应的操作。该系统在存在可能干扰血液成分(如抗坏血酸、丙酮酸、ADP、尿酸和钾离子)时具有稳健性。使用新传感器成功测量了血液中的 ATP 和乳酸水平,回收率良好。使用我们的传感器获得的血液样本的分析结果与使用传统方法获得的结果非常吻合。基于电极的分析和基于微控制器的系统的集成节省了进一步的操作,使在 5 分钟内直接测量 ATP 和乳酸水平成为可能。该传感器可能成为严重传染病管理的有用工具。