| abstract |
The present invention provides methods and kits useful for the detection of genetic alterations and to diagnose a disease condition caused by a genetic alteration. The invention involves two different PCR products, one control 'unaltered' product and one sample product, which may contain 'altered' DNA, that are mixed, denatured, and allowed to re-anneal. The DNA duplexes are then treated enzymatically or chemically to cleave single stranded or mismatched DNA regions resulting from the genetic alterations. The alterations are then quantitated by measuring a decrease in a fluorescent signal resulting from removal of the fluorescent tag. Further embodiments of the invention include a second, different fluorescent tag that is used to measure non-specific degradation of the duplexes. Other embodiments of the invention include the use of avidin coated microbeads to which the duplexes may be attached for quantitation by flow cytometry. Still further embodiments include biological microbeads for this purpose. |