Evaluation of detection performance for single copy of DNA by PCR

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K05494
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 38050:Food sciences-related
• Research Institution: National Agriculture and Food Research Organization
• Principal Investigator: Takabatake Reona 国立研究開発法人農業・食品産業技術総合研究機構, 食品研究部門, 上級研究員 (20463466)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 定量分析 / PCR / 1分子 / DNA

Outline of Final Research Achievements

Generally, it is considered that a single copy of DNA can be detected by PCR, but the hypothesis has not yet been verified. In this study, I attempted to create and utilize a standard molecule that can control the target sequence for the evaluation of the detection performance for a single copy of DNA. To accurately evaluate the detection performance, a standard molecule reliably containing a single copy of the target sequence is indispensable. Then, a single copy of the target sequence and multiple copies of another confirmation sequence were introduced into the same plasmid DNA and used as a standard DNA. The solution containing the standard plasmid was highly diluted, and the presence of the standard plasmid was checked using the confirmation sequence, and the detection performance for a single copy was evaluated using the target sequence.

Free Research Field

食品分析

Academic Significance and Societal Importance of the Research Achievements

化学分析において、偽陰性率とは、「陽性であることが既知の試料を陰性と判定した比率」を表す。しかしながら、従来の分析技術では、偽陰性率に関しては、ほとんどサンプリングに起因するもののみが考慮されてきた。ただし、サンプリングによる偽陰性率は、あくまでも選抜した検査用試料から標的を漏らしてしまう確率を示しており、必ずしも定義通りの偽陰性率の評価にはなっていない。「標的が存在しているにも関わらず検出されなかった」という、真の意味での偽陰性率の評価とは言い難い。本研究によって、PCR検査における真の意味での偽陰性率の評価が実現する。