Development of high-sensitivity detection technology for nucleic acid amplification using metal and metal chelator and verification of its practicality

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K07340
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 52010:General internal medicine-related
• Research Institution: Kyushu University
• Principal Investigator: Hokazono Eisaku 九州大学, 医学研究院, 講師 (60404042)
• Co-Investigator(Kenkyū-buntansha): 内海 健 九州大学, 医学研究院, 教授 (80253798)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 高感度検出 / 核酸増幅 / 金属キレータ / ピロリン酸

Outline of Final Research Achievements

Various amplification techniques have been developed and put to practical use in nucleic acid-based testing. However, the development of technology for detecting the degree of amplification has been mainly based on improved probe detection using fluorescent labeling, and expensive equipment has been indispensable. Therefore, we focused on pyrophosphate, which is produced during nucleic acid amplification, and attempted to develop a simpler technique for detecting the amplified amount of nucleic acids. As a result, we established a high-sensitivity detection method for pyrophosphate and confirmed that the sensitivity of this method is comparable to existing nucleic acid amplification methods. Taking advantage of this simple and rapid detection characteristic, our method is expected to be applied to gene-related tests for disasters including infectious diseases in the future.

Free Research Field

臨床化学

Academic Significance and Societal Importance of the Research Achievements

本検討の成果として、ピロリン酸の高感度検出系の構築に至った。また、既存の核酸増幅技術（PCR法）との比較検討において、本法が核酸の増幅の有無を検出するにあたり遜色ない感度を有していることを確認した。本検討内容はAnalytical Biochemistryへの論文掲載に至った。
本技術は、迅速かつ簡便な新しい核酸増幅定性・定量検出システムとして，緊急時・災害時の電気，水などのライフラインの乏しい医療現場での感染症などの遺伝子検査キッ トの検出系全般への高感度技術の応用展開への可能性を示すものであると考える。