COMPOSITIONS FOR USE IN IDENTIFICATION OF TICK-BORNE PATHOGENS
First Claim
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1. A composition, comprising at least one purified oligonucleotide primer pair that comprises forward and reverse primers, wherein said primer pair comprises nucleic acid sequences that are substantially complementary to nucleic acid sequences of two or more different tick-borne pathogens and/or or endosymbionts, wherein said primer pair is configured to produce amplicons comprising different base compositions that correspond to said two or more different bioagents.
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Abstract
The present invention relates generally to the field of genetic identification and quantification of tick borne pathogens and provides methods, compositions and kits useful for this purpose when combined with molecular mass or base composition analysis.
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30 Claims
- 1. A composition, comprising at least one purified oligonucleotide primer pair that comprises forward and reverse primers, wherein said primer pair comprises nucleic acid sequences that are substantially complementary to nucleic acid sequences of two or more different tick-borne pathogens and/or or endosymbionts, wherein said primer pair is configured to produce amplicons comprising different base compositions that correspond to said two or more different bioagents.
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20. A kit, comprising at least one purified oligonucleotide primer pair that comprises forward and reverse primers that are about 20 to 35 nucleobases in length, and wherein said forward primer comprises at least 70%, at least 80%, at least 90%, at least 95%, or at least 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOS:
- 1-18, and said reverse primer comprises at least 70% sequence identity with a sequence selected from the group consisting of SEQ ID NOS;
19-36.
- 1-18, and said reverse primer comprises at least 70% sequence identity with a sequence selected from the group consisting of SEQ ID NOS;
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21. A method of determining a presence of a tick-borne pathogen and/or or endosymbiont in at least one sample, the method comprising:
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(a) amplifying one or more segments of at least one nucleic acid from said sample using at least one purified oligonucleotide primer pair that comprises forward and reverse primers that are about 20 to 35 nucleobases in length, and wherein said forward primer comprises at least 70%, at least 80%, at least 90%, at least 95%, or at least 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs;
1-18, and said reverse primer comprises at least 70% sequence identity with a sequence selected from the group consisting of SEQ ID NOs;
19-36 to produce at least one amplification product; and(b) detecting said amplification product, thereby determining said presence of said tick-borne pathogen and/or or endosymbiont in said sample. - View Dependent Claims (22, 23, 24)
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26. A system, comprising:
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(a) a mass spectrometer configured to detect one or more molecular masses of amplicons produced using at least one purified oligonucleotide primer pair that comprises forward and reverse primers, wherein said primer pair comprises nucleic acid sequences that are substantially complementary to nucleic acid sequences of two or more different tick-borne pathogen and/or or endosymbiont bioagents; and (b) a controller operably connected to said mass spectrometer, said controller configured to correlate said molecular masses of said amplicons with one or more tick-borne pathogen and/or or endosymbiont bioagent identities. - View Dependent Claims (27, 28, 29, 30)
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Specification