Detection of point mutations in the HBV polymerase gene using a fluorescence intercalator in reverse micelles

Tatsuo Maruyama, Tomoaki Takata, Hirofumi Ichinose, Noriho Kamiya, Hiroyuki Kuma, Naotaka Hamasaki, Hiroyuki Morita, Masahiro Goto

研究成果: ジャーナルへの寄稿記事

7 引用 (Scopus)

抄録

We report a novel and simple method for mutation detection in DNA oligonucleotides using a double-stranded DNA specific dye (SYBR Green I) in nanostructured molecular assemblies, called reverse micelles. The intercalation of SYBR Green I into the duplex DNA exhibits fluorescent emission in a CTAB/isooctane reverse micellar system as well as in an aqueous solution. We found marked differences in the fluorescence intensity between perfectly matched and mismatched 52-mer synthetic oligonucleotides, which were designed to contain the YMDD motif of the hepatitis B virus (HBV) polymerase gene, in a reverse micellar solution. Using this method, we successfully detected a mutation in PCR-amplified oligonucleotides of the HBV polymerase gene in sera of four patients with chronic hepatitis B. This detection method does not require DNA immobilization, chemical modification of DNA, or any special apparatus; it only needs a normal fluorescence spectrophotometer, an inexpensive dye, and just 10 pmol of sample DNA.

元の言語英語
ページ(範囲)575-579
ページ数5
ジャーナルBiotechnology Progress
21
発行部数2
DOI
出版物ステータス出版済み - 3 1 2005

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Intercalating Agents
Micelles
Point Mutation
Hepatitis B virus
Fluorescence
DNA
Oligonucleotides
Genes
Coloring Agents
Mutation
Chronic Hepatitis B
Immobilization
Polymerase Chain Reaction
Serum

All Science Journal Classification (ASJC) codes

  • Biotechnology

これを引用

Detection of point mutations in the HBV polymerase gene using a fluorescence intercalator in reverse micelles. / Maruyama, Tatsuo; Takata, Tomoaki; Ichinose, Hirofumi; Kamiya, Noriho; Kuma, Hiroyuki; Hamasaki, Naotaka; Morita, Hiroyuki; Goto, Masahiro.

:: Biotechnology Progress, 巻 21, 番号 2, 01.03.2005, p. 575-579.

研究成果: ジャーナルへの寄稿記事

Maruyama, Tatsuo ; Takata, Tomoaki ; Ichinose, Hirofumi ; Kamiya, Noriho ; Kuma, Hiroyuki ; Hamasaki, Naotaka ; Morita, Hiroyuki ; Goto, Masahiro. / Detection of point mutations in the HBV polymerase gene using a fluorescence intercalator in reverse micelles. :: Biotechnology Progress. 2005 ; 巻 21, 番号 2. pp. 575-579.
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AU - Ichinose, Hirofumi

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AU - Kuma, Hiroyuki

AU - Hamasaki, Naotaka

AU - Morita, Hiroyuki

AU - Goto, Masahiro

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