Novel high-sensitivity detection method of virus using a hairpin primer PCR

2015 Fiscal Year Final Research Report

• Project/Area Number: 24310165
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Chemical biology
• Research Institution: National Defense Medical College (2014-2015); Osaka University (2012-2013)
• Principal Investigator: Fumie Takei 防衛医科大学校(医学教育部医学科進学課程及び専門課程、動物実験施設、共同利用研究, 医学教育部医学科進学課程, 准教授 (30252711)
• Co-Investigator(Renkei-kenkyūsha): NAKAKTNI Kazuhiko 大阪大学, 産業科学研究所, 教授 (70237303)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Keywords: PCR / 蛍光分子 / 遺伝子検出 / ウイルス検出 / グアニン / シトシンバルジ構造 / RT-PCR

Outline of Final Research Achievements

We have previously described a hairpin primer PCR (HP-PCR) method using a primer having a hairpin tag at the 5′ end and a fluorescent dye (2,7-diamino-1,8-naphthyridine derivative [DANP]) bound to the cytosine bulge (C-bulge) embedded in the hairpin-tag. It was turn-off type of system. In this research, we tried to develop the high-sensitivity detection method using a HP-PCR and it applied to detect some kinds of virus.
In this research, we developed the new type of PCR monitoring system using DANP bound to the primers as a covalent and in which the fluorescent signal increases as PCR progresses. This was a turn-on of PCR using fluorescently and modified DNA.

Free Research Field

ゲノム化学