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METHOD FOR DETECTING TUBERCULOSIS COMPLEX-DERIVED DNA

Foreign code F170009088
File No. (S2016-0104-N0)
Posted date May 30, 2017
Country WIPO
International application number 2016JP082390
International publication number WO 2017077999
Date of international filing Nov 1, 2016
Date of international publication May 11, 2017
Priority data
  • P2015-218146 (Nov 6, 2015) JP
Title METHOD FOR DETECTING TUBERCULOSIS COMPLEX-DERIVED DNA
Abstract As a result of in-depth research of primary and probe designs, digital PCR conditions, and the like with which the various bacterial strains of tuberculosis complex can be detected in order to establish a method for detecting tuberculosis complex-derived DNA in plasma by digital PCR, the inventors of the present invention established a method with which tuberculosis complex-derived DNA can be detected from the plasma of a tuberculosis patient on a level of several copies/20 µL reaction solution by digital PCR using a specific primer and probe targeting IS6110 and gyrB.
Scope of claims [claim1]
1. The forward primer of the base arrangement which is shown in arrangement number 1, the reverse primer of the base arrangement which is shown in arrangement number 2, and the probe of the base arrangement which is shown in arrangement number 3 are included, the digital PCR primer and probe set for tubercle bacillus group IS6110 detection.
[claim2]
2. The forward primer of the base arrangement which is shown in arrangement number 4, the reverse primer of the base arrangement which is shown in arrangement number 5, and the probe of the base arrangement which is shown in arrangement number 6 are included, the digital PCR primer and probe set for tubercle bacillus group gyrB detection.
[claim3]
3. DNA which is extracted is designated as the mold from the blood sample which is separated from the suffering inspection person, the fact that it does digital PCR the primer and probe set of statement making use of each of them at least is included in claim 1 and 2, method of detection of tubercle bacillus group origin DNA.
[claim4]
4. The aforementioned blood sample is the blood plasma or the serum, method of claim 3 statement.
[claim5]
5. Use density of each primer is 700nM-1100nM, use density of the probe is 100nM-150nM, claim method of 3 or 4 statements.
[claim6]
6. It does digital PCR the DNA standard which is DNA of the direct condition which includes the part territory of the tubercle bacillus group gene which includes the expansion object domain of the digital PCR primer as a mold, furthermore it includes the fact that threshold for distinguishing the positive areole is decided with the cluster analysis of signal strength of each areole of the said DNA standard, claim either 3 or 5 in 1 sections method of statement.
[claim7]
7. DNA standard for IS6110 detection type includes the base arrangement which is shown in arrangement number 11, DNA standard for gyrB detection type includes the base arrangement which is shown in arrangement number 12, method of claim 6 statement.
[claim8]
8. It is done in order to assist the decision of the curative effect of the monitor of the condition of detection, the tubercle bacillus group infection symptom or the tubercular symptom of tubercle bacillus group infection, appraisal or the tubercle bacillus group infection symptom or the tubercular symptom of tubercular symptom emergence risk, claim either 3 or 7 in 1 sections method of statement.
[claim9]
9. The primer and probe set of statement each of them is included at least in claim 1 and 2, the tubercle bacillus group origin DNA detection reagent or the kit for digital PCR.
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • YOKOHAMA CITY UNIVERSITY
  • Inventor
  • YAMAMOTO MASAKI
  • USHIO RYOTA
IPC(International Patent Classification)
Specified countries (WO201777999)
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 DJ 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 KW 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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