Balanced translocation in comparative hybridization

• US 8,076,074 B2
• Filed: 11/29/2005
• Issued: 12/13/2011
• Est. Priority Date: 11/29/2005
• Status: Expired due to Fees

• Current Assignee: Quest Diagnostics Investments Incorporated (Quest Diagnostics Incorporated)
• Original Assignee: Quest Diagnostics Investments Incorporated (Quest Diagnostics Incorporated)
• Inventors: Mohammed, Mansoor S.
• Primary Examiner(s): POHNERT, STEVEN C
• Application Number:
  US11/288,982
• Publication Number:
  US 20070122820A1
• Time in Patent Office: 2,205 Days
• Field of Search: None
• US Class Current: 435/6.1
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574

Abstract

The present invention provides comparative genomic hybridization methods for detecting and mapping chromosomal or genetic abnormalities associated with various diseases or with predisposition to various diseases, or to detecting the phenomena of large scale copy number variants. The method includes hybridization with one or more probes for detecting balanced translocations. Such probes may be complementary to the moving genomic segment which is translocated or may be complementary to the translocation break point.

Claims

1. A method of detecting the presence or absence of chromosomal abnormalities and a balanced translocation in a test sample of genomic nucleic acid, comprising:

(ii) hybridizing to a genomic nucleic acid array;

a) said test sample of genomic nucleic acid;

b) a reference genomic nucleic acid;

(ii) hybridizing at least one probe for detecting a balanced translocation to said test and reference genomic nucleic acid hybridized to said array, wherein at least one of said probes has a sequence that spans a translocation break point; and

(iii) determining the relative amount of hybridized test and reference nucleic acids hybridized to the array as well as determining hybridization of said at least one probe for detecting a balanced translocation to said test and reference genomic nucleic acid hybridized to said array, wherein detecting differential binding to said array of said test sample compared to said reference genomic nucleic acid indicates the presence of a chromosomal abnormality, and wherein detecting differential binding of at least one probe for detecting a balanced translocation to said test sample compared to said reference genomic nucleic acid indicates the presence of a balanced translocation.

2. The method of claim 1 wherein said test and reference genomic nucleic acids are labeled with different detectable labels and said at least one probe for detecting a translocation is labeled with a detectable label.

3. The method of claim 1 wherein multiple probes for detecting multiple translocations are used.

4. The method of claim 3 wherein said multiple probes for multiple translocations are each detectably labeled with labels that individually distinguish each of the translocation probes.

5. The method of claim 4 wherein said test and reference genomic nucleic acids are labeled with labels that are distinguishable from the labels used for the probes for detecting a translocation.

6. The method of claim 1 wherein said test and reference nucleic acids are labeled with the same detectable label and said at least one probe for a translocation is labeled with a detectable label different from the said label for the test and reference sample.

7. The method of claim 6 wherein the amount of hybridized test and reference nucleic acids are determined by:

a) determining a signal for the detectable label hybridized to the array representing the total of hybridized test and reference nucleic acid;

b) treating the hybridized nucleic acids to selectively remove said detectable label from one of said test or reference nucleic acids, wherein said detectable label is linked to the test and reference nucleic acids through a first and second chemical linkage, respectively, and wherein one of said first linkage or said second linkage is susceptible to the treatment step;

c) determining a signal for the detectable label hybridized to the array following step b), which represents one of said hybridized test or reference nucleic acid; and

d) determining a signal for the other of the hybridized test or reference by using the signal from c) and b).

8. The method of claim 6 wherein the amount of hybridized test and reference nucleic acids are determined by:

a) determining a signal representing one of the test or reference nucleic acid hybridized to the array by contacting the hybridized array with a first complex comprising a detectable label and a first entity, wherein said first complex selectively reacts with one of said test or reference nucleic acid via a first tag attached to said test or reference nucleic acid, said first entity and tag being members of a specific binding pair;

b) determining a signal representing the total of the hybridized test and reference nucleic acid by contacting the hybridized array of step a) with a second complex comprising the detectable label and a second entity, wherein said second complex selectively reacts with the other of said test or reference nucleic acid via a second tag attached to the other of said test or reference nucleic acid detected in step b), said second entity and second tag being members of a specific binding pair; and

c) using the signal from a) and b) to obtain the signal for the other of said hybridized test or reference nucleic acid not detected in step a).

9. The method of any of claims 8, wherein said first tag and said second tag each comprises a unique oligonucleotide capture sequence.

10. The method of claim 9, wherein said first complex and said second complex each comprises an oligonucleotide complementary to one of said unique oligonucleotide capture sequences.

11. The method of claim 9, wherein said first complex and said second complex each comprises an oligonucleotide complementary to one of said unique oligonucleotide capture sequences in a dendrimeric construct.

12. The method of claim 1 wherein the amount of hybridized test and reference nucleic acids are determined by:

a) determining a signal representing one of the test or reference nucleic acid hybridized to the array, wherein said detectable label has been attached before hybridization;

b) determining a signal representing the total of the hybridized test and reference nucleic acid by contacting the hybridized array with a first complex comprising a detectable label and an entity, wherein said first complex selectively reacts with the other of said test or reference nucleic acid via a tag attached to said other of the test or reference nucleic acid;

c) using the signal from a) and b) to obtain the signal for the other of said hybridized test or reference nucleic acid not detected in step a).

13. The method of any of claims 12, wherein said tag comprises an oligonucleotide capture sequence.

14. The method of claim 13, wherein said first complex comprises an oligonucleotide complementary to said oligonucleotide capture sequence.

15. The method of claim 1 wherein said nucleic acid array comprises a plurality of nucleic acid segments each contained within an artificial chromosome.

16. The method of claim 15, wherein said artificial chromosome is a bacterial artificial chromosome (BAC).

17. The method of claim 1 wherein said nucleic acid array comprises a plurality of nucleic acid segments, each nucleic acid segment of said plurality being between about 1,000 and about 1,000,000 nucleotides in length.

18. The method of claim 1, wherein said nucleic acid array comprises a plurality of nucleic acid segments that collectively comprise the nucleic acid sequence of at least one chromosome.

19. The method of claim 17, wherein said plurality of nucleic acid segments comprise segments of chromosomal sequence of about 1-4 megabases in length.

20. The method of claim 17, wherein said plurality of nucleic acid segments collectively comprise the nucleic acid sequence of all chromosomes of a cell.

21. The method of claim 1, wherein said nucleic acid array comprises a plurality of nucleic acid segments that collectively comprise at least about 50 genomic regions of interest.

22. The method of claim 1, wherein said nucleic acid array comprises at least about 300 discrete locations where a nucleic acid of the plurality of nucleic acid segments is present.

23. The method of claim 1, wherein said nucleic acid array comprises at least about 500 discrete locations where a nucleic acid of the plurality of nucleic acid segments is present.

24. The method of claim 1, wherein at least one detectable label is a fluorophore.

25. The method of claim 1, wherein said test sample is obtained from a patient.

26. The method of claim 1, wherein said test nucleic acids and reference nucleic acids are genomic nucleic acids.

27. The method of claim 1, wherein said probe comprises a detectable label comprising a dendrimer complex with an oligonucleotide capture sequence complimentary to an unbound region of a probe partially bound to the array.

28. The method of claim 27, wherein the oligonucleotide region for the probe for translocations is at least 12 by in length.

29. The method of claim 27, wherein the oligonucleotide region for the probe for translocation is at least 15 bp in length.

30. The method of claim 27, wherein the oligonucleotide region for the probe for translocation is at least 18 bp in length.