Development of nucleic acid extraction-free microbial analysis technology that can be used rapidly onsite

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K04328
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 22060:Environmental systems for civil engineering-related
• Research Institution: Nagaoka National College of Technology
• Principal Investigator: Kawakami Shuji 長岡工業高等専門学校, 環境都市工学科, 准教授 (00610461)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 環境微生物 / aptamer / シングルセル

Outline of Final Research Achievements

With the aim of developing a technique for detecting microorganisms without the need for DNA extraction, this study examined the results of real-time PCR measurements of the number of aptamers bound to bacterial samples prepared by step dilution and with different numbers of bacteria present. The experimental results showed that there was a correlation between the initial bacterial abundance and the amount of aptamer, indicating the possibility of quantifying the number of bacteria. In addition, all samples showed a faster amplification rise compared to the amplification curve when targeting conventional genes, indicating the possibility of detection with higher sensitivity than conventional gene quantification methods

Free Research Field

環境微生物

Academic Significance and Societal Importance of the Research Achievements

特定の微生物を定量する方法としてreal-time PCR法があるが, real-time PCR法は， DNAの数を計測できる技術であり，標的とする微生物のDNAを抽出後，標的遺伝子をPCR法で増幅させ，その増幅の速度からもとの試料中の微生物数を算出する手法である．しかしながら，DNAの抽出に専門的な知識や，コンタミネーションを防ぐクリーンな実験室の施設を必要とすることから，屋外等の現場レベルでの作業が困難という問題点がある．本研究はDNA抽出不要の微生物定量技術を提案するものであり、これら課題を解決できる可能性がある。