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CLAMPING PROBE

Foreign code F160008826
File No. (S2015-0122-N0)
Posted date Aug 10, 2016
Country WIPO
International application number 2015JP081403
International publication number WO 2016072516
Date of international filing Nov 6, 2015
Date of international publication May 12, 2016
Priority data
  • P2014-226332 (Nov 6, 2014) JP
Title CLAMPING PROBE
Abstract Developed and provided is a method whereby a mutated gene that exists in a gene pool together with a number of wild type genes can be easily, economically and highly sensitively detected. Provided is a clamping probe that is capable of binding to target nucleic acid molecules at two regions, i.e., first and second nucleic acid complementary regions, and thus distinguishing these target nucleic acid molecules depending on a difference in complementarity to a wild type target nucleic acid molecule and complementarity to a mutated target nucleic acid molecule.
Scope of claims [claim1]
1. The target nuclear acid molecule 1 or being the clamping probe in order to detect the known mutation with the substitution, deficiency or addition of several bases,
The aforementioned clamping probe the 1st target nuclear acid complimentary territory, to consist of the one chain nuclear acid molecule which includes the 2nd target nuclear acid complimentary territory, the hairpin territory, and two these chain territories, at the same time
3' end of the aforementioned 1st target nuclear acid complimentary territory and 5' end of 2nd target nuclear acid complimentary territory, or
3' end of the aforementioned 2nd target nuclear acid complimentary territory and 5' end of 1st target nuclear acid complimentary territory
In each case the hairpin territory is connected on the one hand, at the same time two these chain territories are connected on the other hand and become,
Here,
The aforementioned 1st target nuclear acid complimentary territory consists of complimentary base arrangement in base arrangement of the 1st target nuclear acid territory,
It consists of the 15-30 base to which the aforementioned 1st target nuclear acid territory includes the aforementioned known mutation region in the wild type target nuclear acid molecule and continues,
The aforementioned 2nd target nuclear acid complimentary territory consists of complimentary base arrangement in base arrangement of the 2nd target nuclear acid territory,
The aforementioned 2nd target nuclear acid territory consists of the 15-30 base which 5' terminal side of the aforementioned 1st target nuclear acid territory or 3' is adjacent to terminal side on the target nuclear acid molecule and is located,
As for the aforementioned hairpin territory,
Two these chain parts which consist of the 3-10 base the complimentary base arrangement mutually and,
Either of said two these chain parts 1 sets 5' end and 3' the one chain part which consists of the base arrangement of the 3-10 base which connects end
To consist of, and
The aforementioned two these chain territories consist of the 3-10 base the complimentary base arrangement mutually
The aforementioned clamping probe.
[claim2]
2. The aforementioned 1st target nuclear acid complimentary territory base arrangement and includes the mismatch region of the 1-3 base which compliment of the 1st target nuclear acid territory is not done, in claim 1 the clamping probe of statement.
[claim3]
3. The aforementioned 1st target nuclear acid complimentary territory and/or between 2nd target nuclear acid complimentary territory and hairpin territory, and/or
The aforementioned 1st target nuclear acid complimentary territory and/or between 2nd target nuclear acid complimentary territory and two these chain territories
The spacer territory which consists of the base arrangement of the 1-5 base which lies between is included, in claim 1 or 2 the clamping probe of statement.
[claim4]
4. Separation 5 of the aforementioned two these chain territories' end or separation 3' end the G quartet territory was connected at least on the one hand, either of the claim 1-3 in one section the clamping probe of statement.
[claim5]
5. It is the KRas gene where the aforementioned target nuclear acid molecule shows with SEQ ID NO 1, either of the claim 1-4 in one section the clamping probe of statement.
[claim6]
6. The aforementioned mutation, it is the substitution mutation at least in one which is selected from the group which consists of 34 rank, 35 rank, and 38 rank base, in claim 5 the clamping probe of statement.
[claim7]
7. It is the EGFR gene where the aforementioned target nuclear acid molecule shows with SEQ ID NO 2, either of the claim 1-4 in one section the clamping probe of statement.
[claim8]
8. The aforementioned mutation, in each case is selected from the group which consists of 2115 rank, 2573 rank and 2582 rank it is the substitution mutation in one base, in claim 7 the clamping probe of statement.
[claim9]
9. Either of the claim 1-4 mixing the clamping probe of statement into one section in the nuclear acid sample which includes the target nuclear acid molecule, the target nuclear acid molecule and the process which makes the clamping probe connect,
The process which detects the mutation of the target nuclear acid molecule on the basis of the difference of binding power of the target nuclear acid molecule and the clamping probe due to the presence of mutation of the target nuclear acid molecule
The method of detecting the presence of known mutation on the target nuclear acid molecule which is included.
[claim10]
10. Being the detection with the quantitative difference of the expansion product the detection which is based on the difference of the aforementioned binding power due to nuclear acid expansion method,
The 1st target nuclear acid territory of the target nuclear acid molecule and the territory where the 2nd target nuclear acid territory is included is expanded with the aforementioned nuclear acid expansion method, in claim 9 method of statement.
[claim11]
11. It is the detection the detection which is based on the difference of the aforementioned binding power by the molecular sieve, in claim 9 method of statement.
[claim12]
12. It is the KRas gene where the aforementioned target nuclear acid molecule shows with SEQ ID NO 1, either of the claim 9-11 in one section method of statement.
[claim13]
13. The aforementioned mutation, it is the substitution mutation at least in one which is selected from the group which consists of 34 rank, 35 rank, and 38 rank base, in claim 12 method of statement.
[claim14]
14. It is the EGFR gene where the aforementioned target nuclear acid molecule shows with SEQ ID NO 2, either of the claim 9-11 in one section method of statement.
[claim15]
15. The aforementioned mutation, in each case is selected from the group which consists of 2115 rank, 2573 rank and 2582 rank it is the substitution mutation in one base, in claim 14 method of statement.
[claim16]
16. Being affection distinction method of the colon cancer,
The process which manufactures the nuclear acid sample from the organism sample which is obtained from the suffering *** body,
The process which detects the presence of known mutation on the KRas gene of statement in claim 12 or 13 making use of the nuclear acid sample which is obtained,
When the KRas gene which possesses mutation in the nuclear acid sample is detected, as for the suffering *** body when you have contracted a disease in the colon cancer, the process which is distinguished
The aforementioned method of including.
[claim17]
17. Being affection distinction method of the non small cell lung cancer,
The process which manufactures the nuclear acid sample from the organism sample which is obtained from the suffering *** body,
The process which detects the presence of known mutation on the EGFR gene of statement in claim 14 or 15 making use of the nuclear acid sample which is obtained,
When the EGFR gene which possesses mutation in the nuclear acid sample is detected, as for the suffering *** body when you have contracted a disease in the non small cell lung cancer, the process which is distinguished
The aforementioned method of including.
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • OSAKA CITY UNIVERSITY
  • Inventor
  • TACHIBANA AKIRA
  • TANABE TOSHIZUMI
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG

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