RU2010151495A - Композиции и способы определения хромосомных аберраций с новыми буферами для гибридизации - Google Patents
Композиции и способы определения хромосомных аберраций с новыми буферами для гибридизации Download PDFInfo
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Abstract
1. Способ гибридизации in situ для определения по меньшей мере одного молекулярного зонда и определения присутствия хромосомной аберрации в хромосомной ДНК, включающий ! - получение по меньшей мере одного молекулярного зонда, который гибридизуется с мишенью в хромосомной ДНК, ! - получение хромосомной ДНК, ! - получение композиции для гибридизации, содержащей по меньшей мере один полярный апротонный растворитель в количестве, эффективном для денатурации двунитевых нуклеотидных последовательностей, где полярный апротонный растворитель не является диметилсульфоксидом (ДМСО), ! - объединение молекулярного зонда, хромосомной ДНК и композиции для гибридизации в течение по меньшей мере периода времени, достаточного для гибридизации молекулярного зонда с мишенью, и ! - определение мишени. ! 2. Способ по п.1, где указанная композиция для гибридизации содержит по меньшей мере один молекулярный зонд. ! 3. Способ по п.1, где полярный апротонный растворитель определен согласно любому из пп.7-20. ! 4. Способ по любому из пп.1-3, где стадия гибридизации занимает менее 8 ч, менее 1 часа, менее 30 минут. ! 5. Способ диагностики врожденного генетического расстройства, рака или инфекции, связанных с хромосомной аберрацией, включающий ! - получение образца ткани от субъекта, где образец ткани содержит нуклеиново-кислотную последовательность, ! - определение, присутствует ли хромосомная аберрация в нуклеиново-кислотной последовательности, в соответствии со способом по любому из пп.1-4, и ! - диагностику врожденного генетического расстройства, рака или инфекции, если хромосомная аберрация присутствует в образце ткани. ! 6. Применение композици�
Claims (21)
1. Способ гибридизации in situ для определения по меньшей мере одного молекулярного зонда и определения присутствия хромосомной аберрации в хромосомной ДНК, включающий
- получение по меньшей мере одного молекулярного зонда, который гибридизуется с мишенью в хромосомной ДНК,
- получение хромосомной ДНК,
- получение композиции для гибридизации, содержащей по меньшей мере один полярный апротонный растворитель в количестве, эффективном для денатурации двунитевых нуклеотидных последовательностей, где полярный апротонный растворитель не является диметилсульфоксидом (ДМСО),
- объединение молекулярного зонда, хромосомной ДНК и композиции для гибридизации в течение по меньшей мере периода времени, достаточного для гибридизации молекулярного зонда с мишенью, и
- определение мишени.
2. Способ по п.1, где указанная композиция для гибридизации содержит по меньшей мере один молекулярный зонд.
3. Способ по п.1, где полярный апротонный растворитель определен согласно любому из пп.7-20.
4. Способ по любому из пп.1-3, где стадия гибридизации занимает менее 8 ч, менее 1 часа, менее 30 минут.
5. Способ диагностики врожденного генетического расстройства, рака или инфекции, связанных с хромосомной аберрацией, включающий
- получение образца ткани от субъекта, где образец ткани содержит нуклеиново-кислотную последовательность,
- определение, присутствует ли хромосомная аберрация в нуклеиново-кислотной последовательности, в соответствии со способом по любому из пп.1-4, и
- диагностику врожденного генетического расстройства, рака или инфекции, если хромосомная аберрация присутствует в образце ткани.
6. Применение композиции, содержащей молекулярный зонд, который определяет нуклеотидную последовательность, связанную с хромосомной аберрацией, и композицию для гибридизации, содержащую от 1% (об/об) до 95% (об/об) по меньшей мере одного полярного апротонного растворителя по любому из пп.7-20, в гибридизационном анализе на определение нуклеотидной последовательности, связанной с хромосомной аберрацией.
7. Композиция для гибридизации, содержащая:
(а) первый молекулярный зонд, который определяет нуклеотидную последовательность, связанную с хромосомной аберрацией,
(б) по меньшей мере один полярный апротонный растворитель, имеющий циклическую основную структуру, в количестве, эффективном для денатурации двунитевых нуклеотидных последовательностей, и
(в) раствор для гибридизации,
где полярный апротонный растворитель не является диметилсульфоксидом (ДМСО).
8. Композиция для гибридизации по п.7, где хромосомная аберрация представляет собой анеуплоидию, потенциальную точку разрыва, инсерцию, инверсию, делецию, дупликацию, амплификацию гена, перестройку или транслокацию.
9. Композиция для гибридизации по п.7, где первый молекулярный зонд определяет ALK, BCL2, BCL3, BCL6, BCL10, BCL12, BCR, CCND1, Е2А, EGFR, ETV6, FIP1L1, HER2, IGH, IGK, IGL, MALT1, MLL (ALL-1, HTRX1, HRX), MYC (с-Мус), РАХ5, PDGFRA, PDGFRB, SIL, TCF3 (Е2А, ITF1), TCL1A, TCRAD, TCRB, TCRG, теломер, TLX1, TLX3 (HOX11L2, RNX) или ТОР2А.
10. Композиция для гибридизации по п.7, где первый молекулярный зонд определяет хромосомную аберрацию, выбранную из t(1;14) (q34;q11), t(1;19) (q23;p13), t(2;5), t(2;18) (q12;q21), t(2;8), t(4;11), t(4;11) (q21;q23), t(6;11) (q27;q23), t(7;22) (p22;q12), t(8;14), t(8;22), t(9;11) (p22;q23), t(9;22) (q34;q11), t(10;14) (q24;q11), t(11;14), t(11;14) (p13;q11), t(11;19) (q23;p13), t(14;18) (q23;q21), t(14;18), t(18;22) (q21;q11) и t(21;22)(q22;q12).
11. Композиция для гибридизации по п.7, дополнительно содержащая второй молекулярный зонд, причем предпочтительно второй молекулярный зонд определяет референсную последовательность.
12. Композиция для гибридизации по п.7, где первый молекулярный зонд и второй молекулярный зонд определяют последовательности, фланкирующие одну или более чем одну из потенциальных точек разрыва, или находящиеся в ее пределах.
13. Композиция для гибридизации по п.7, где молекулярный зонд дополнительно содержит метку.
14. Композиция для гибридизации по п.7, где концентрация полярного апротонного растворителя в композиции для гибридизации находится в интервале от 1% (об/об) до 95% (об/об), либо от 10% (об/об) до 20% (об/об).
15. Композиция для гибридизации по п.7, где полярный апротонный растворитель имеет лактоновую, сульфоновую, сульфитную, нитрильную и/или карбонатную функциональную группу.
16. Композиция для гибридизации по п.7, где полярный апротонный растворитель имеет дисперсионный параметр растворимости в интервале от 17,7 МПа1/2 до 22,0 МПа1/2, полярный параметр растворимости в интервале от 13 МПа1/2 до 23 МПа1/2 и параметр растворимости за счет водородных связей в интервале от 3 МПа1/2 до 13 МПа1/2.
17. Композиция для гибридизации по п.7, где полярный апротонный растворитель выбран из группы, состоящей из:
где Х представляет собой О и R1 представляет собой алкилдиил, и
где Х является необязательным и, если присутствует, выбран из О или S;
где Z является необязательным и, если присутствует, выбран из О или S;
где А и В независимо представляют собой О или N или S или часть алкилдиила или первичного амина;
где R представляет собой алкилдиил; и где Y представляет собой О или S или С.
19. Композиция для гибридизации по п.7, содержащая по меньшей мере один полярный апротонный растворитель, декстрансульфат и буфер.
20. Композиция для гибридизации по п.7, содержащая 15% по меньшей мере одного полярного апротонного растворителя, 20% декстрансульфата, 600 мМ хлорида натрия, 10 мМ цитратного буфера рН 6,2.
21. Набор, содержащий:
(а) первый молекулярный зонд, который определяет нуклеотидную последовательность, связанную с хромосомной аберрацией, и
(б) композицию для гибридизации, содержащую по меньшей мере один полярный апротонный растворитель по любому из пп.7-20 в количестве, эффективном для денатурации двунитевых нуклеотидных последовательностей,
где полярный апротонный растворитель не является диметилсульфоксидом (ДМСО).
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US5608908P | 2008-05-27 | 2008-05-27 | |
US61/056.089 | 2008-05-27 | ||
DKPA200800727 | 2008-05-27 | ||
GB61/056.089 | 2008-05-27 | ||
DKPA200800727 | 2008-05-27 | ||
US15568309P | 2009-02-26 | 2009-02-26 | |
US61/155,683 | 2009-02-26 | ||
DKPA200900278 | 2009-02-27 | ||
DKPA200900278 | 2009-02-27 | ||
PCT/IB2009/006548 WO2009147537A2 (en) | 2008-05-27 | 2009-05-27 | Compositions and methods for detection of chromosomal aberrations with novel hybridization buffers |
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CN (3) | CN102084004B (ru) |
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