Detection of nucleic acid sequence differences using the ligase detection reaction with addressable arrays
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C07H-021/04
C12N-015/11
출원번호
US-0072442
(2011-03-25)
등록번호
US-8288521
(2012-10-16)
발명자
/ 주소
Barany, Francis
Barany, George
Hammer, Robert P.
Kempe, Maria
Blok, Herman
Zirvi, Monib
출원인 / 주소
Cornell Research Foundation, Inc.
대리인 / 주소
LeClairRyan, a Professional Corporation
인용정보
피인용 횟수 :
3인용 특허 :
114
초록▼
The present invention describes a method for identifying one or more of a plurality of sequences differing by one or more single base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences. The method includes a ligation phase, a capture phase, and a detectio
The present invention describes a method for identifying one or more of a plurality of sequences differing by one or more single base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences. The method includes a ligation phase, a capture phase, and a detection phase. The ligation phase utilizes a ligation detection reaction between one oligonucleotide probe, which has a target sequence-specific portion and an addressable array-specific portion, and a second oligonucleotide probe, having a target sequence-specific portion and a detectable label. After the ligation phase, the capture phase is carried out by hybridizing the ligated oligonucleotide probes to a solid support with an array of immobilized capture oligonucleotides at least some of which are complementary to the addressable array-specific portion. Following completion of the capture phase, a detection phase is carried out to detect the labels of ligated oligonucleotide probes hybridized to the solid support.
대표청구항▼
1. A kit for identifying one or more of a plurality of sequences differing by single-base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences comprising: a ligase;a collection of oligonucleotide probe sets, each set characterized by (a) a first oligonucleo
1. A kit for identifying one or more of a plurality of sequences differing by single-base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences comprising: a ligase;a collection of oligonucleotide probe sets, each set characterized by (a) a first oligonucleotide probe comprising a target-specific portion and (b) a second oligonucleotide probe comprising a target-specific portion and a further portion, wherein the further portion comprises a nucleic acid sequence of greater than sixteen nucleotides that differs for each different target-specific portion, and wherein the nucleic acid sequence of a complement to one further portion differs from the nucleic acid sequence of a complement to another further portion in the collection by at least 25%, when aligned to each other. 2. The kit according to claim 1 further comprising a solid support anda collection of capture oligonucleotides immobilized at particular sites on the solid support, wherein the capture oligonucleotides have nucleotide sequences complementary to the further portions and, wherein the solid support and the capture oligonucleotides form an addressable array. 3. The kit according to claim 2, wherein the solid support is made from a material selected from the group consisting of plastic, ceramic, metal, resin, gel, glass, silicon, and composites thereof. 4. The kit according to claim 2, wherein the solid support is in a form selected from the group consisting of beads, spheres, plates, strands, slides, discs, membranes, films, and composites thereof. 5. The kit according to claim 4, wherein the solid support comprises a plurality of beads, wherein each bead comprises one or more types of capture oligonucleotides from the collection of capture oligonucleotides. 6. The kit according to claim 4, wherein the solid support comprises a plurality of spheres, wherein each sphere comprises one or more types of capture oligonucleotides from the collection of capture oligonucleotides. 7. The kit according to claim 2, wherein the solid support has an array of positions with a plurality of capture oligonucleotides attached to the array of positions. 8. The kit of claim 2, wherein the solid support comprises a high-density array of capture oligonucleotides immobilized thereto. 9. The kit according to claim 2, wherein the collection of capture oligonucleotides comprises at least 25 different capture oligonucleotides. 10. The kit according to claim 9, wherein the collection of capture oligonucleotides comprises at least 1000 different capture oligonucleotides. 11. The kit according to claim 9, wherein the collection of capture oligonucleotides comprises 25 to 1200 different capture oligonucleotides. 12. The kit according to claim 1 further comprising: amplification primers suitable for amplification of the target nucleotide sequences anda polymerase. 13. The kit according to claim 2, wherein each capture oligonucleotide in the collection of capture oligonucleotides comprises a nucleotide sequence of greater than sixteen nucleotides. 14. The kit according to claim 13, wherein each capture oligonucleotide in the collection comprises a nucleotide sequence of 24 nucleotides or less. 15. The kit according to claim 1, wherein the collection of oligonucleotide probe sets comprises at least 25 different oligonucleotide probe sets. 16. The kit according to claim 15, wherein the collection of oligonucleotide probe sets comprises at least 1000 different oligonucleotide probe sets. 17. The kit according to claim 15, wherein the collection of oligonucleotide probe sets comprises 25 to 1200 different oligonucleotide probe sets. 18. A kit for identifying one or more of a plurality of target nucleotide sequences in a sample comprising: a ligase;a plurality of oligonucleotide probe sets, each set characterized by (a) a first oligonucleotide probe, having a target-specific portion and an address-specific portion and (b) a second oligonucleotide probe, having a target-specific portion; anda collection of capture oligonucleotides wherein each type of capture oligonucleotide in the collection comprises a nucleotide sequence complementary to an address-specific portion, wherein the address-specific portion is comprised of a nucleotide sequence which is distinct from that of the target-specific portions, and wherein each type of capture oligonucleotide in the collection hybridizes to its complement under uniform hybridization conditions but differs by at least 25% in nucleotide sequence, when aligned to another type of capture oligonucleotide in the collection. 19. The kit according to claim 18 further comprising: amplification primers suitable for amplification of the target nucleotide sequences anda polymerase. 20. The kit according to claim 18, wherein the address-specific portion of the first oligonucleotide probe comprises a nucleotide sequence of greater than sixteen nucleotides. 21. The kit according to claim 18, wherein each capture oligonucleotide in the collection of capture oligonucleotides comprises a nucleotide sequence of greater than sixteen nucleotides. 22. The kit according to claim 21, wherein each capture oligonucleotide in the collection of capture oligonucleotides comprises a nucleotide sequence of 24 nucleotides or less. 23. The kit according to claim 18 further comprising: a solid support, wherein capture oligonucleotides of the collection are immobilized at particular sites on the solid support. 24. The kit according to claim 23, wherein the collection of capture oligonucleotides and the solid support form an addressable array. 25. The kit according to claim 23, wherein the solid support comprises a plurality of beads, wherein each bead comprises one or more types of capture oligonucleotides from the collection. 26. The kit according to claim 23, wherein the solid support comprises a plurality of spheres, wherein each sphere comprises one or more types of capture oligonucleotides from the collection. 27. The kit according to claim 18, wherein the plurality of oligonucleotide probe sets comprises at least 25 different oligonucleotide probe sets. 28. The kit according to claim 27, wherein the plurality of oligonucleotide probe sets comprises at least 1000 different oligonucleotide probe sets. 29. The kit according to claim 27, wherein the plurality of oligonucleotide probe sets comprises 25 to 1200 different oligonucleotide probe sets. 30. The kit according to claim 18, wherein the collection of capture oligonucleotides comprises at least 25 different capture oligonucleotides. 31. The kit according to claim 30, wherein the collection of capture oligonucleotides comprises at least 1000 different capture oligonucleotides. 32. The kit according to claim 30, wherein the collection of capture oligonucleotides comprises 25 to 1200 different capture oligonucleotides.
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