Title:
METHOD FOR DETECTION OF MUTATED POLYNUCLEOTIDE
Document Type and Number:
WIPO Patent Application WO/2012/002447
Kind Code:
A1
Abstract:
Disclosed is a highly sensitive genetic mutation detection method which can get information on genetic mutation from a CTC.
Specifically disclosed is a method for detecting a mutated polynucleotide in a circulating tumor cell, which is characterized by comprising the following steps: step 1: a polynucleotide solution preparation step of extracting a cell sample containing 1 to 10000 circulating tumor cells with an aqueous solvent containing proteinase K to produce a polynucleotide solution; step 2: an amplification step of subjecting a reaction solution that contains all of the polynucleotide solution produced in step 1 to the gene amplification by a PCR method to selectively amplify a detection target region in the mutated polynucleotide which has a likelihood of being contained in the polynucleotide solution; and step 3: a detection step of subjecting (1) an amplification product produced in step 2 and (2) a reaction solution containing a probe that is an LNA comprising a nucleotide sequence absolutely complementary to the detection target region in the mutated polynucleotide to the gene amplification by a PCR method to selectively amplify the detection target region in the mutated polynucleotide, thereby detecting the mutated polynucleotide.
Inventors:
MATSUMOTO HIDEO (JP)
MIYAMOTO SAKU (JP)
MIYAMOTO SAKU (JP)
Application Number:
PCT/JP2011/064937
Publication Date:
January 05, 2012
Filing Date:
June 29, 2011
Export Citation:
Assignee:
TAKEDA PHARMACEUTICAL (JP)
MATSUMOTO HIDEO (JP)
MIYAMOTO SAKU (JP)
MATSUMOTO HIDEO (JP)
MIYAMOTO SAKU (JP)
International Classes:
C12Q1/68; C12N15/09; C12Q1/02
Foreign References:
| JP2009544283A | 2009-12-17 | |||
| JP2008538282A | 2008-10-23 |
Other References:
MIHO UHARA ET AL.: "Idenshi Hen'i Tokuiteki PCR-ho o Mochiita Junkan Shuyo Saibo no Kenshutsuho no Kaihatsu", THE JAPANESE JOURNAL OF CLINICAL PATHOLOGY, vol. 57, 2009, pages 232, 0 - 285
NAGAI Y. ET AL.: "Genetic heterogeneity of the epidermal growth factor receptor in non-small cell lung cancer cell lines revealed by a rapid and sensitive detection system, the peptide nucleic acid-locked nucleic acid PCR clamp.", CANCER RES., vol. 65, no. 16, 2005, pages 7276 - 7282
KAZUTOSHI ISOBE ET AL.: "Ketsuekichu no Junkan Shuyo Saibo (CTC) o Mochiita EGFR mutation Kenshutsu no Kento", THE JOURNAL OF THE JAPAN SOCIETY FOR CANCER THERAPY, vol. 45, no. 2, 21 September 2010 (2010-09-21), pages 1023
KAZUHIRO KITAMURA ET AL.: "PNA-LNA PCR clamp-ho Oyobi PCR-invader-ho ni yoru EGFR Idenshi Hen'i Kaiseki no validation Shiken", THE JOURNAL OF THE JAPAN SOCIETY FOR CANCER THERAPY, vol. 45, no. 2, 21 September 2010 (2010-09-21), pages 1023
NEZAS A. ET AL.: "Detection of circulating tumor cells in bladder cancer patients.", CANCER TREAT. REV., vol. 35, no. 3, 2009, pages 272 - 279
NAGAI Y. ET AL.: "Genetic heterogeneity of the epidermal growth factor receptor in non-small cell lung cancer cell lines revealed by a rapid and sensitive detection system, the peptide nucleic acid-locked nucleic acid PCR clamp.", CANCER RES., vol. 65, no. 16, 2005, pages 7276 - 7282
KAZUTOSHI ISOBE ET AL.: "Ketsuekichu no Junkan Shuyo Saibo (CTC) o Mochiita EGFR mutation Kenshutsu no Kento", THE JOURNAL OF THE JAPAN SOCIETY FOR CANCER THERAPY, vol. 45, no. 2, 21 September 2010 (2010-09-21), pages 1023
KAZUHIRO KITAMURA ET AL.: "PNA-LNA PCR clamp-ho Oyobi PCR-invader-ho ni yoru EGFR Idenshi Hen'i Kaiseki no validation Shiken", THE JOURNAL OF THE JAPAN SOCIETY FOR CANCER THERAPY, vol. 45, no. 2, 21 September 2010 (2010-09-21), pages 1023
NEZAS A. ET AL.: "Detection of circulating tumor cells in bladder cancer patients.", CANCER TREAT. REV., vol. 35, no. 3, 2009, pages 272 - 279
Attorney, Agent or Firm:
SAEGUSA & PARTNERS (JP)
Patent business corporation 3 Edakuni [Hajime] patent firm (JP)
Patent business corporation 3 Edakuni [Hajime] patent firm (JP)
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