| abstract |
The invention is directed towards a method of amplifying target nucleic acids by using two oligonucleotide probe complement pairs. Each member of the probe pair contains a chemical functionality group which permits linkage of the probes when the functionality groups are adjacent to one another following hybridization of the probe pairs to the template. One probe in each pair is composed of two regions, a first region which hybridizes to the target and which contains the chemical functionality group and a second, protecting region which prevents target independent joining as shown in the figure. The other probe in the pair contains the corresponding chemical functionality group. Upon joining of a first probe pair, an amplification can proceed in which the newly joined first probe pair can serve as a template for the second, complementary probe pair, which can in turn serve as a template for unjoined first probe pairs. This cyclic amplification is sensitive enough for a discriminative amplification of sequences which differ by merely a point mutation and therefore is suitable for point mutation detection and genotype determination as well as for the determination of the presence or absence of a specific nucleic acid in a sample. |