FIELD: medicine.
SUBSTANCE: method involves producing an amplification reaction mixture containing at least one target sequence from nucleic acid, a primer of said sequence, a hybridisation probe of said sequence, a DNA fluorescent dye and reagents required for amplification. Said target sequence from nucleic acid is amplified with the help of said process of asymmetric amplification of nucleic acids to form one-strand and two-strand amplicons. Then two-strand amplicons are detected by the DNA fluorescent dye, and at least one said one-strand amplicon is detected by fluorophore marked hybridisation probe at temperature below an annealing point of the primer of the target sequence. It is followed by normalising the fluorescent signal specific for the sequence in the form of the calculated relation of fluorescence of said one-strand product to fluorescence of said two-strand amplicons.
EFFECT: presented invention enables low-variation homogenous detection of the one-strand amplification product of the process of asymmetric amplification of nucleic acids.
9 cl, 24 dwg, 16 ex
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Authors
Dates
2012-09-10—Published
2005-10-17—Filed