| CPC C12Q 1/689 (2013.01) [C12Q 1/6895 (2013.01)] | 27 Claims |
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1. A method of identifying one or more specific microbial species in a sample from a subject, the method comprising:
depleting eukaryotic DNA from the sample;
lysing one or more microbial cells in the sample, wherein the lysing of one or more microbial cells releases a plurality of microbial genetic materials;
isolating the plurality of microbial genetic materials;
amplifying the plurality of microbial genetic materials;
incubating the amplified microbial genetic materials with a plurality of DNA Invading Artificial Nucleic Acids (DIANAs) for less than 10 minutes at a temperature of below 65° C.; and
detecting binding of one or more of the plurality of DIANAs to the amplified microbial genetic material of its respective single species or group of microbes, wherein the detection of binding indicates the presence of one or more specific microbial species or groups of microbes in the sample,
wherein:
the amplified microbial genetic materials are invaded by at least one of the DIANAs in the plurality of DIANAs during the incubation,
one or more of the plurality of the DIANAs is a peptide nucleic acid, a locked nucleic acid, and/or a bridged nucleic acid, and
the peptide nucleic acid, locked nucleic acid, and/or bridged nucleic acid:
comprises a sequence that shares at least 60% identity with a sequence selected from the group consisting of SEQ ID NOS: 20-571,
comprises the complement, reverse, or reverse complement of a sequence that shares at least 60% identity with a sequence selected from the group consisting of SEQ ID NOS: 20-571,
comprises a sequence selected from the group consisting of SEQ ID NOS: 20-571 that lacks six or fewer bases at either or both ends, and/or
comprises the complement, reverse, or reverse complement of a sequence selected from the group consisting of SEQ ID NOS: 20-571 that lacks six or fewer bases at either or both ends.
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