Title:
METHYLCYTOSINE DETECTION METHOD
Document Type and Number:
WIPO Patent Application WO/2014/142228
Kind Code:
A1
Abstract:
Provided is a method for detecting methylcytosine using an anti-methylcytosine antibody, wherein the methylation of specific cytosine in genome DNA methylation is selectively detected, and the quantitative properties and reliability thereof are improved. A method for detecting the methylated state of cytosine at a specific position contained within nucleic acid, said method comprising: a step in which the nucleic acid is fragmented by a restriction enzyme; a step in which a double-strand nucleic acid is formed between the fragmented nucleic acid and a single-strand nucleic acid having a solid-phase bond part and base sequence which hybridize with the fragmented nucleic acid, but which do not form a base pair with cytosine at a specific position within the fragmented nucleic acid; a step in which the double-strand nucleic acid is bonded on a solid phase using the solid-phase bond part; and a step in which the quantity of antibodies bonded to the the double-strand nucleic acid on the solid phase is measured. As a result of this configuration, everything apart from the cytosine being measured forms base pairs with guanine, and is not detected by the antibodies, and it is possible to thereby selectively detect the presence of methylation of specific cytosine of the base sequence to be measured from a large genome.
Inventors:
KURITA RYOJI (JP)
YANAGISAWA HIROYUKI (JP)
YOSHIOKA KYOKO (JP)
NIWA OSAMU (JP)
YANAGISAWA HIROYUKI (JP)
YOSHIOKA KYOKO (JP)
NIWA OSAMU (JP)
Application Number:
PCT/JP2014/056633
Publication Date:
September 18, 2014
Filing Date:
March 13, 2014
Export Citation:
Assignee:
NAT INST OF ADVANCED IND SCIEN (JP)
International Classes:
C12N15/09; C12Q1/44; C12Q1/68; G01N33/53
Domestic Patent References:
| WO2005080565A1 | 2005-09-01 |
Foreign References:
| JP2012230019A | 2012-11-22 |
Other References:
KURITA R. ET AL.: "DNA methylation analysis triggered by bulge specific immuno-recognition.", ANAL. CHEM., vol. 84, no. 17, 2012, pages 7533 - 7538, XP055213627, DOI: doi:10.1021/ac301702y
RYOJI KURITA ET AL.: "DNA Bulge Tokuiteki na Kotai Ninshiki o Riyo shita Cytosine no Methyl- ka Shindan", THE JAPAN SOCIETY FOR ANALYTICAL CHEMISTRY DAI 61 NENKAI KOEN YOSHISHU, 2012, pages 51
RYOJI KURITA ET AL.: "DNA Bulge ni Haichi sareta Methyl Cytosine to Kotaikan deno Sogo Sayo Kaiseki", DAI 73 KAI ABSTRACTS OF THE SYMPOSIUM OF THE JAPAN SOCIETY FOR ANALYTICAL CHEMISTRY, May 2013 (2013-05-01), pages 10
RYOJI KURITA ET AL.: "DNA Bulge o Riyo shita Ichi Sentakuteki Methyl Cytosine Keisoku Chip no Kaihatsu", SOCIETY FOR CHEMISTRY AND MICRO- NANO SYSTEMS DAI 27 KAI KENKYUKAI KOEN YOSHISHU, May 2013 (2013-05-01), pages 17
RYOJI KURITA ET AL.: "DNA Bulge Tokuiteki na Kotai Ninshiki o Riyo shita Cytosine no Methyl- ka Shindan", THE JAPAN SOCIETY FOR ANALYTICAL CHEMISTRY DAI 61 NENKAI KOEN YOSHISHU, 2012, pages 51
RYOJI KURITA ET AL.: "DNA Bulge ni Haichi sareta Methyl Cytosine to Kotaikan deno Sogo Sayo Kaiseki", DAI 73 KAI ABSTRACTS OF THE SYMPOSIUM OF THE JAPAN SOCIETY FOR ANALYTICAL CHEMISTRY, May 2013 (2013-05-01), pages 10
RYOJI KURITA ET AL.: "DNA Bulge o Riyo shita Ichi Sentakuteki Methyl Cytosine Keisoku Chip no Kaihatsu", SOCIETY FOR CHEMISTRY AND MICRO- NANO SYSTEMS DAI 27 KAI KENKYUKAI KOEN YOSHISHU, May 2013 (2013-05-01), pages 17
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