WO2014058355A8 - Способ количественного анализа терминальных нуклеотидов g-цепи теломерной днк человека - Google Patents

Способ количественного анализа терминальных нуклеотидов g-цепи теломерной днк человека Download PDF

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WO2014058355A8
WO2014058355A8 PCT/RU2013/000892 RU2013000892W WO2014058355A8 WO 2014058355 A8 WO2014058355 A8 WO 2014058355A8 RU 2013000892 W RU2013000892 W RU 2013000892W WO 2014058355 A8 WO2014058355 A8 WO 2014058355A8
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dna
chain
overhangs
telomeric
terminal
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PCT/RU2013/000892
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English (en)
French (fr)
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WO2014058355A1 (ru
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Елена Андреевна ЧИРЯСОВА
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Елена Андреевна ЧИРЯСОВА
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Application filed by Елена Андреевна ЧИРЯСОВА filed Critical Елена Андреевна ЧИРЯСОВА
Priority to EP13846152.0A priority Critical patent/EP2907872B1/en
Publication of WO2014058355A1 publication Critical patent/WO2014058355A1/ru
Publication of WO2014058355A8 publication Critical patent/WO2014058355A8/ru

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6851—Quantitative amplification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Analytical Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Pathology (AREA)
  • Hospice & Palliative Care (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Изобретение предназначено для выявления изменений нуклеотидных окончаний ДНК теломерных областей хромосом,. происходящих в ходе клеточного цикла. Способ основывается на новом подходе без использования процедуры лигирования. Способ включает этапы предварительной экстракции геномной ДНК, выделения фракции однонитевых G-оверхенгов теломерной ДНК и последующую амплификацию их минусовой цепи с дуплекс-специфическим анализом. Этапы амплификации включают модификацию 3'-концов теломерных G-оверхенгов с помощью терминальной дезоксинуклеотидилтрансферазы и осуществляются с использованием набора праймеров, представленного в последовательностях SEQ ID NO:1-5. Дуплекс-специфический анализ основан на гибридизации минусовых цепей оверхенгов с набором специфических флуоресцентно-меченных зондов (SEQ ID NO:6-11), соответствующих шести возможным вариантам нуклеотидных окончаний G-цепи теломерной ДНК человека. Степень разгорания зондов в присутствии фермента дуплекс-специфической нуклеазы служит критерием присутствия соответствующего варианта нуклеотидного окончания ДНК и мерой его количественного содержания. Полученные профили процентного содержания терминальных триплетов G-цепи теломерной ДНК служат пролиферативными маркерами, указывающими на степень активации деления клетки и определяющими временную продолжительность клеточного цикла. Изобретение обеспечивает повышение чувствительности и точности количественного анализа терминальных нуклеотидов по сравнению с методами, основанными на лигировании. Способ позволяет получать диагностические результаты в иммунологии, онкологии, трансплантологии, косметологии, дерматологии, геронтологии, клеточной биотехнологии и других областях, заинтересованных в оценке пролиферативного статуса клеток и методах его направленного регулирования.
PCT/RU2013/000892 2012-10-10 2013-10-09 Способ количественного анализа терминальных нуклеотидов g-цепи теломерной днк человека WO2014058355A1 (ru)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP13846152.0A EP2907872B1 (en) 2012-10-10 2013-10-09 Methods and compositions for the quantitive analysis of terminal nucleotides of a g chain of human telomeric dna

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
RU2012143263 2012-10-10
RU2012143263/10A RU2508407C9 (ru) 2012-10-10 2012-10-10 Способ количественной оценки терминальных нуклеотидов g-цепи теломерной днк человека с помощью полимеразной цепной реакции и дуплекс-специфического анализа, наборы синтетических олигонуклеотидных праймеров и зондов для осуществления этого способа

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WO2014058355A1 WO2014058355A1 (ru) 2014-04-17
WO2014058355A8 true WO2014058355A8 (ru) 2017-08-17

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PCT/RU2013/000892 WO2014058355A1 (ru) 2012-10-10 2013-10-09 Способ количественного анализа терминальных нуклеотидов g-цепи теломерной днк человека

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EP (1) EP2907872B1 (ru)
RU (1) RU2508407C9 (ru)
WO (1) WO2014058355A1 (ru)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106834545A (zh) * 2017-03-13 2017-06-13 苏州市立医院 高危人乳头瘤病毒试剂盒及检测方法
US10604794B2 (en) 2017-05-10 2020-03-31 The Board Of Regents Of The University Of Texas System Method to measure the shortest telomeres

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5741677A (en) * 1995-06-07 1998-04-21 Geron Corporation Methods for measuring telomere length
ZA200108461B (en) * 2000-10-27 2002-06-06 Celanese Chem Europe Gmbh Process of telomerizing conjugated dienes.
US20040265815A1 (en) * 2001-06-23 2004-12-30 Baird Duncan Martin Method for determination of telomere length
US20090306182A1 (en) * 2002-02-20 2009-12-10 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF MAP KINASE GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
CA2695414A1 (en) * 2007-08-10 2009-02-19 Tina Holding Aps Method for estimating telomere length
RU2434940C1 (ru) * 2010-07-20 2011-11-27 Александр Борисович Смолянинов Способ измерения длины теломер клеток лейкоконцентрата пуповинной крови

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RU2012143263A (ru) 2013-02-20
RU2508407C2 (ru) 2014-02-27
EP2907872A4 (en) 2016-02-24
EP2907872B1 (en) 2017-12-20
RU2508407C9 (ru) 2014-05-20
WO2014058355A1 (ru) 2014-04-17
EP2907872A1 (en) 2015-08-19

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