2022年 新着論文 8 神経変性病態学分野、生体防御病態代謝研究分野から論文が発表されました

Electrochemical enzyme-based blood ATP and lactate sensor for a rapid and straightforward evaluation of illness severity

Biosens Bioelectron. 2022 Feb 15;198:113832. doi: 10.1016/j.bios.2021.113832. Epub 2021 Nov 23.

Authors

Keine Nishiyama  1 Ryohei Mizukami  1 Shizuka Kuki  1 Akihiko Ishida  2 Junji Chida  3 Hiroshi Kido  4 Masatoshi Maeki  5 Hirofumi Tani  5 Manabu Tokeshi  6

Affiliations

  • 1 Graduate School of Chemical Sciences and Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo, 060-8628, Japan.
  • 2 Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo, 060-8628, Japan. Electronic address: ishida-a@eng.hokudai.ac.jp.
  • 3 Division of Molecular Neurobiology, Institute of Advanced Medical Sciences, Tokushima University, Kuramoto-cho 3-18-15, Tokushima, 770-8503, Japan.
  • 4 Division of Pathology and Metabolome Research for Host Defense, Institute of Advanced Medical Sciences, Tokushima University, Kuramoto-cho 3-18-15, Tokushima, 770-8503, Japan.
  • 5 Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo, 060-8628, Japan.
  • 6 Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo, 060-8628, Japan; Institute of Innovation for Future Society, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8601, Japan; Innovative Research Center for Preventive Medical Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8601, Japan.

Abstract

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.

Keywords: ATP; Blood analysis; Integrated electrode; Lactate; Prussian Blue.

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