Development of a new molecular diagnostic technique to detect 20 HAB species within 15 min

2016 Fiscal Year Final Research Report

• Project/Area Number: 26550085
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Environmental risk control and evaluation
• Research Institution: Fisheries Research and Education Agency
• Principal Investigator: Nagai Satoshi 国立研究開発法人水産研究・教育機構, 中央水産研究所, グループ長 (80371962)
• Research Collaborator: MIYAMOTO Nobuhiko 株式会社カネカ, 研究部探索グループ, 研究主任
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 核酸クロマト / multiplex-PCR / 有毒渦鞭毛藻 / 有害赤潮 / 分子同定

Outline of Final Research Achievements

In this study, the Kaneka DNA chromatography chip (KDCC) for Alexandrium species was successfully developed that enables the simultaneous detection of five Alexandrium species. This method is utilized a DNA-DNA hybridization technology. This technique is simple, because it is possible to detect the target species by naked eye soon after (< 5 min) applying two μL of PCR amplicon and 65 μL of development buffer to the sample pad of the chip. Further, this technique is relatively inexpensive (does not require expensive laboratory equipment such as real-time Q-PCR machines or DNA microarray detectors, but a thermal cycler). Regarding the detection limit of KDCC for the five Alexandrium species, it varied among species and they were <0.1-10 pg and equivalent to 5-500 copies of rRNA genes, indicating that the technique is sensitive enough for practical use to detect several cells of the target species from 1 L of seawater.

Free Research Field

分子同定