Id Biomedical Corporation

Peter D Duck

Faouzi Bekkaoui

William L Crosby

Lynn P Cloney

Zora D Modrusan

Richard H Tullis

John McNevin

Isabelle A Piche

Alfred C Wong

A method for detecting a target nucleic acid molecule is provided, comprising the steps of (a) reacting a mixture comprising (i) a target nucleic acid molecule; (ii) a single-stranded nucleic acid probe containing a scissile linkage; (iii) an enzyme capable of cleaving the probe portion of a double-stranded target-probe complex at the scissile linkage; and (iv) ribosomal protein and/or spermine, under conditions and for a time sufficient to allow the target nucleic acid and probe to hybridize to each other and form a double-stranded target-probe complex, followed by cleavage of the probe and cycling of the target to a new uncleaved probe, such that one or more portions of the cleaved nucleic acid probe are released from the target-probe complex; and (b) determining whether cleaved portions of the nucleic acid probe are produced, and thereby detecting the presence of the target nucleic acid.

Additives for use in cycling probe reactions

Briefly stated, the present invention provides novel compositions and methods for detecting target nucleic acid sequences utilizing adjacent sequence-enzyme molecules. Within one aspect of the present invention, oligonucleotide-enzyme fusion molecules are provided, comprising an enzyme capable of cleaving scissile linkages and an oligonucleotide having the structure (NA.sub.1).sub.x --S.sub.z --(NA.sub.2).sub.y wherein NA.sub.1 and NA.sub.2 are nucleic acid sequences, S is a scissile nucleic acid linkage, x, y and z are integers from 1 to 100 and n is an integer from 1 to 10.

Compositions and methods for detecting target nucleic acid sequences

A method for determining whether a selected nucleic acid molecule differs by one or more nucleotides from a desired sequence (the desired sequence may be defined to be either a native (e.g., wild type) or non-native (e.g., mutant) nucleotide sequence). The potential difference between the selected nucleic acid molecule and the desired nucleic acid sequence may be due to a germ or somatic mutation and may be, for example, a substitution, deletion addition or transversion of one or more nucleotides. Thus, for example, the method is useful for detection of mutations in oncogenes or mutations in other genes of interest, such as the transmembrane conductance regulator gene, which is implicated in cystic fibrosis.

Cycling probe cleavage detection of nucleic acid sequences

Growing community of inventors

Burnaby, Canada

Peter D Duck