MULTIPLEX DETECTION

摘要

The present invention relates to a method for designing polynucleotide primer/probes capable of specific and mismatch tolerant hybridization useful in multiplex detection and comprising capture and amplification steps. Said method comprises identifying a nucleation site of 6-9 nucleotides, and building a nucleotide sequence by positioning nucleotides substantially complementary to said nucleation site and positioning nucleotides outside said site and introducing either universal base analogues such as deoxyinosine at sites of variation and/or positioning degenerate nucleotide positions at sites of variation thereby obtaining a continous primer/probe. The present invention also relates multiplex detection of nucleic acids and primer/probes and kits for use in such a method, as well as a computer program for determining how the primer/probes should be constructed for an optimised result.