Patent ID: 6190868
Filing Date: 2001-02-20
Classification: C12N,C12Q

Abstract:
A method for identifying one or more nucleic acids in a sample comprising a plurality of nucleic acids having different nucleotide sequences, said method comprising:(a) providing a sample contacted by one or more recognition means, each of said recognition means recognizing a different target nucleotide sequence or different set of target nucleotide subsequences;(b) generating one or more signals from said sample by amplifying one or more sequences in said sample with at least one amplification means, each generated signal arising from a nucleic acid in said sample and comprising a representation of (i) the size of the sequence comprising said target nucleotide subsequences in said nucleic acid, and (ii) the identities of said target subsequences in said nucleic acid;(c) searching a nucleotide sequence database for sequences that have a corresponding size and identified target subsequence as one or more of the nucleic acids generating said signal, said database comprising a plurality of known nucleotide sequences of nucleic acids that may be present in the sample, a sequence from said database matching the generated signal when the sequence from said database has both (i) the same size of the sequence comprising said target nucleotide subsequences as is represented by the generated signal, and (ii) the same target subsequences as are represented by the generated signal, or target subsequences that are members of the same sets of target subsequences represented by the generated signal, thereby identifying a putative matching sequence;(d) contacting under amplifying conditions said sample with said amplification means and one or more poisoning primers to form a reamplified signal corresponding to said putative matching sequence, wherein said poisoning primers are capable of hybridizing to and amplifying only nucleic acids having said putative matching sequence, said poisoning primers being unlabeled or differentially labeled from said amplification means and having a sequence comprising a first subsequence that is a portion of said putative matching sequence and a second subsequence; and(e) comparing the reamplified signal of said putative matching sequence to the signal of said putative matching sequence in step (b), wherein a decrease in the reamplified signal relative to the amplified signal verifies the identity of said matching sequences thereby identifying one or more nucleic acids in said sample.