装置不要!生物汚染をわずか15分かつ室温下で目視検出──核酸アプタマー誘導型ハイブリダイゼーション連鎖反応を利用した機器不要なATPの迅速目視検出法の開発に成功──

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2026-02-16 東京大学

東京大学大学院総合文化研究科の吉本敬太郎准教授らとダイキン工業の共同研究グループは、核酸アプタマー誘導型ハイブリダイゼーション連鎖反応(HCR)と金ナノ粒子(AuNP)の比色反応を最適化し、ATPを室温・約15分で目視検出できる機器不要の手法を開発した。ssDNA濃度やMgCl₂濃度など律速因子を体系的に解析し、反応速度と感度のトレードオフを解消。AuNPの局在表面プラズモン共鳴を利用し、ATP存在下で赤色から青紫色への明確な色変化を実現した。加温不要・迅速・高特異的なオンサイト検査法として、食品・医薬品製造現場の衛生管理や微生物汚染スクリーニングへの応用が期待される。成果は『Analytical Methods』に掲載。

装置不要!生物汚染をわずか15分かつ室温下で目視検出──核酸アプタマー誘導型ハイブリダイゼーション連鎖反応を利用した機器不要なATPの迅速目視検出法の開発に成功──
本研究で開発した室温下15分で目視検出可能なATP検出の概要

<関連情報>

金ナノ粒子を用いた迅速な視覚的ATP検出のためのアプタマー誘導ハイブリダイゼーション連鎖反応の最適化 Optimization of aptamer-triggered hybridization chain reaction for rapid visual ATP detection using gold nanoparticles

Shengli Zhou,Hiroko Fukaya,Shunsuke Watanuki,Wei Liu,Maasa Yokomori,Muneyuki Matsuo,Kazuya Okada,Yukina Yoshioka and Keitaro Yoshimoto
Analytical Methods  Published:11 Feb 2026
DOI:https://doi.org/10.1039/D5AY01738F

Abstract

The development of fast, simple, and visual methods for detecting adenosine triphosphate (ATP) is crucial for point-of-care diagnostics and environmental monitoring. Colorimetric assays based on the integration of DNA aptamer-triggered hybridization chain reaction (HCR) with gold nanoparticles (AuNPs) offer high potential, but existing methods often suffer from prolonged detection times (e.g., >130 min). To address this limitation, this study systematically investigated and optimized the core parameters of the aptamer-triggered HCR system and its subsequent mixing conditions with AuNPs to achieve a rapid visual detection platform. The HCR kinetics were accelerated by the high concentrations of single-stranded DNA (ssDNA) components, particularly the H0 initiator, as well as by the critical presence of divalent magnesium ions (Mg2+), which also support the functionality of the aptamer. This optimization allowed for a degree of HCR progression comparable to the traditional 24-hour incubation to be achieved within just 60 minutes. Subsequently, recognizing the inherent trade-off between the high ssDNA concentration required for fast HCR and the AuNP dispersion stability, we optimized the mixing ratio between the HCR product and the AuNP solution. Under these optimized conditions, the assay demonstrated the capability for rapid visual detection of ATP at the 100 µM level within a total assay time of 15 minutes, representing a significant acceleration compared to previously reported methods. Further improvements in sensitivity are anticipated through future fine-tuning of AuNP parameters and the use of post-reaction salt aggregation enhancers.

生物化学工学
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