Method for detecting allelic imbalance

• Main Authors: Fox, Jayne C, Haque, Kemal, Little, Stephen
• Format: Patent
• Language: English
• Published: 01.07.2003

This invention relates to a diagnostic method for detecting genetic changes associated, inter alia, with the development of cancer. The method detects allelic imbalance (AI), such as loss of heterozygosity (LOH), in nucleic acid from an individual and can be applied to samples in which only a small proportion of the cells display AI, thereby allowing the early identification of mutations which lead to the progression of cancer. The invention also relates to amplification primers for use in the method and to diagnostic kits containing them. A method for detecting allelic imbalance (AI) in sample nucleic acid, which method comprises providing multiple copies of a target nucleic acid region present in the sample, the target nucleic acid region comprising a marker of heterozygosity, separating the multiple copies into individual strands then allowing the individual strands to reanneal under conditions which permit the formation of homoduplexes and heteroduplexes, removing any heteroduplexes so formed, subjecting the remaining homoduplexes to the above steps of separation, reannealing and heteroduplex removal one or more times so that any difference in the initial ratio of allelic variants is amplified, and detecting the presence or absence of AI by reference to any difference in allele ratio so detected.