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2017 Fiscal Year Final Research Report

Development of a highly sensitive RNA virus detection method by artificial nucleoproteins

Research Project

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Project/Area Number 15H05229
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section海外学術
Research Field Medical genome science
Research InstitutionOsaka University

Principal Investigator

KAIHATSU KUNIHIRO  大阪大学, 産業科学研究所, 特任准教授(常勤) (70419464)

Co-Investigator(Kenkyū-buntansha) 黒須 剛  国立感染症研究所, ウイルス第一部, 主任研究官 (70432432)
Project Period (FY) 2015-04-01 – 2018-03-31
Keywordsウイルス / バイオテクノロジー / 遺伝子 / 核酸 / 蛋白質
Outline of Final Research Achievements

At Ministry of Health of Thailand, we analyzed blood sample of dengue infected patients and succeeded to identify (1) the viral serotypes (1-4 type) on the virus infected cultured cells using fluorescent labeled antibodies. Then, (2) EDTA was added to the blood supernatant, and genomic RNA was identified after the viral RNA isolation, double stranded DNA amplification by PCR. (3)dengue virus specific IgM and IgG were detected by anti-IgM and anti-IgG on a plate and detected by ELISA method.
Based on the above described method, viral gene and the protein complex was labeled by chemically-modified antibody. The complex was captured by peptide nucleic acid on the test line in the sequence specific manner. It allow us to identify the presence of dengue virus in the specimen.

Free Research Field

核酸化学

Academic Significance and Societal Importance of the Research Achievements

ヒトは、蚊が媒介するデングウイルスに複数回感染すると、デングショック熱やデング出血熱など重篤な感染症状を発症する。従来の抗体診断キットを用いた場合、デングウイルスに複数回感染した検体中の抗原検出感度が低下することがわかった。一方で、本プロジェクトで開発したペプチド核酸を用いれば同ウイルスの遺伝子/蛋白質複合体をその場で検出できることが明らかとなった。これらの知見はデング熱感染重症化の早期処置に有用となる新しい診断手法になりうると期待する。

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Published: 2019-03-29   Modified: 2020-03-30  

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