METHOD FOR COMPETITIVE ALLELE-SPECIFIC CDNA SYNTHESIS AND DIFFERENTIAL AMPLIFICATION OF THE CDNA PRODUCTS

摘要

The present invention provides a method for the detection of the presence of RNA variants comprising the step of performing a competitive cDNA synthesis comprising a first primer specific to a first RNA variant, a second primer specific to a second RNA variant, an RNA-dependent DNA polymerase, and RNA from said sample as a template, wherein said first primer and said second primer comprise an allele-specific nucleotide portion, a target-specific sequence and tag units with a common sequence and/or a discriminating sequence so that the sequence of said tag units is not complementary to said first or second RNA variant, wherein each of the cDNA products obtained from said competitive cDNA synthesis consists of the sequence of only one primer extended by the sequence complementary to one of the target RNA variants.

主权项

1. Method for the detection of the presence of RNA variants in a sample containing RNA, which is suspected to comprise one or more RNA variants of interest, the method comprising the steps of:
a) performing a competitive cDNA synthesis comprising a first primer specific to a first RNA variant, a second primer specific to a second RNA variant, an RNA-dependent DNA polymerase, and RNA from said sample as a template, wherein said first primer and said second primer comprise an allele-specific nucleotide portion, a target-specific sequence and tag units with a common sequence and/or a discriminating sequence so that the sequence of said tag units is not complementary to said first or second RNA variant, wherein each of the cDNA products obtained from said competitive cDNA synthesis consists of the sequence of only one primer extended by the sequence complementary to one of the target RNA variants; b) performing an amplification reaction so that at least part of the cDNA synthesis products obtained from step a) are amplified; c) detecting the presence of amplification reaction products corresponding to the first and/or second RNA variant obtained from step b) utilizing the presence of the allele-specific nucleotide portion or the discriminating sequence of said tag units in said amplification reaction products.