METHOD FOR DETECTING A RECOMBINANT EVENT

• US 20120015369A1
• Filed: 07/18/2011
• Published: 01/19/2012
• Est. Priority Date: 08/27/2004
• Status: Active Grant

• Current Assignee: Esoterix Genetic Laboratories LLC (Laboratory Corporation Of America Holdings)
• Original Assignee: Esoterix Genetic Laboratories LLC (Laboratory Corporation Of America Holdings)
• Inventors: Shuber, Anthony P.
• Granted Patent: US 8,389,220 B2
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material

Abstract

Methods relating to isolating and amplifying chimeric nucleic acid molecules are provided. The methods of the invention are useful for detecting chromosome translocation events associated with diseases or conditions, such as cancer.

Claims

1. A method of amplifying a portion of a chimeric nucleic acid in a heterogeneous sample, the method comprising the steps of

exposing a sample comprising a first nucleic acid, a second nucleic acid, and a chimeric nucleic acid containing a portion of said first nucleic acid and a portion of said second nucleic acid to a capture probe capable of hybridizing to said portion of said first nucleic acid, thereby to capture said chimeric nucleic acid if present in said sample;

removing uncaptured nucleic acid from said sample; and

amplifying, from said chimeric nucleic acid, said portion of said second nucleic acid.

2. The method of claim 1, wherein the presence of an amplified chimeric nucleic acid is indicative of disease.

3. The method of claim 1, wherein said chimeric nucleic acid is a recombinant nucleic acid.

4. The method of claim 1, wherein said chimeric nucleic acid is RNA.

5. The method of claim 4, wherein RNA in said sample is purified prior to said exposing step.

6. The method of claim 4, further comprising the step of preparing a cDNA from said RNA.

7. The method of claim 1, wherein said amplifying step selectively amplifies said portion of said second nucleic acid present in said chimeric nucleic acid.

8. The method of claim 7, wherein said amplifying step comprises exposing said captured nucleic acid to a first primer that is complementary to a portion of said first nucleic acid and a second primer that is complementary to said second nucleic acid.

9. A method for detecting a chromosome translocation event in a DNA sample comprising

adding a DNA sample to an immobilized DNA capture probe capable of hybridizing to said DNA sample, wherein said DNA sample is repeatedly exposed to the immobilized DNA capture probe using reverse-field electrophoresis;

removing the captured DNA sample from the DNA capture probe and

amplifying the captured DNA sample.

10. The method of claim 9, wherein the DNA capture probe is immobilized on one or more beads.

11. The method of claim 9, wherein the DNA capture probe is immobilized on a gel associated with a membrane.

12. The method of claim 9, wherein two or more DNA capture probes are immobilized.

13. The method of claim 9, wherein the captured DNA sample is amplified using a PCR reaction.

14. The method of claim 13, wherein a first primer and a second primer are present in the PCR reaction.

15. The method of claim 14, wherein the first primer corresponds to a region of a first chromosome.

16. The method of claim 14, wherein the second primer corresponds to a region of a second chromosome.