Method for detecting a recombinant event
View Patent ↗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.
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.