FIELD: chemistry.
SUBSTANCE: invention relates to biotechnology, namely to a method of detecting a nucleic acid target-sequence from DNA or a mixture of nucleic acids with the application of a target signal generating primer (TSG-primer). The method includes hybridisation of the nucleic acid target-sequence with the TSG-primer, which contains a hybridisable nucleotide sequence, complementary to the nucleic acid target-sequence, and a reporter molecule and a quencher molecule. The TSG-primer does not represent a loop-hairpin structure. In case if the TSG-primer does not hybridise with the nucleic acid target-sequence, the reporter molecule and the quencher molecule are located close to each other spatially without forming loop-hairpin structures, which makes it possible for the quencher molecule to quench the signal from the reporter molecule. In case the primer hybridises with the nucleic acid target-sequence, the reporter molecule and the quencher molecule are separated spatially, making it not possible for the quencher molecule to quench the signal from the reporter molecule, which results in the generation and detection of the signal indicating the presence of the nucleic acid target-sequence. Bringing the product in contact with a matrix polymerase of nucleic acids under conditions of primer elongation in such a way that the reaction of 3′-elongation is induced on 3′-end of the TSG-primer. Detection of the signal, indicating the presence of the nucleic acid target-sequence.
EFFECT: claimed invention makes it possible to detect the nucleic acid target-sequence with high sensitivity and specificity.
21 cl, 21 dwg, 7 ex
