Patent ID: 6512105
Filing Date: 2003-01-28
Classification: C12Q,Y02A

Abstract:
A method for making a probe for use in a hybridization assay which comprises constructing a oligonucleotide that is sufficiently complementary to hybridize to a region of rRNA, or the encoding DNA, selected to distinguish one or more non-viral target species from at least one non-viral non-target species, said oligonucleotide comprising a target-complementary sequence obtained by:a) aligning rRNA sequences, or the encoding DNA sequences, of said target species and said non-target species to identify a variable region; and b) designing said target-complementary sequence of said oligonucleotide by substantially maximizing the complementarity of said oligonucleotide to said variable region present in said target species while substantially minimizing the complementarity of said oligonucleotide to said variable region present in said non-target species, such that a duplex formed between said oligonucleotide and said target species has a higher Tm than a duplex formed between said oligonucleotide and said non-target species, wherein said variable region is located in an rRNA sequence or DNA sequence corresponding to a target region selected from the group consisting of: bases 60-100 of E. coli 16S rRNA or the encoding DNA; and bases 405-480 of E. coli 16S rRNA or the encoding DNA, and wherein the percent similarity in the rRNA sequences of said non-target species and said target species is between 90 and 99%.