PNA oligomers, oligomer sets, methods and kits pertaining to the detection of
View Patent ↗This invention is related to novel PNA probes, probe sets, methods and kits pertaining to the determination of Bacillus anthracis.
1. A PNA oligomer comprising a probing nucleobase sequence that is at least ninety percent homologous to a nucleobase sequence, or its complement, selected from the group consisting of: CCA-SSG-GTA-TCD-DTC (Seq. ID No. 1) and TTC-AAA-GGC-TCC-CGC (Seq. ID No. 2).
2. The PNA oligomer of claim 1 , wherein the probing nucleobase sequence is one hundred percent homologous to one of Seq. ID No. 1 or Seq. ID No. 2, or their complements.
3. The PNA oligomer of claim 1 , wherein the oligomer is unlabeled.
4. The PNA oligomer of claim 1 , wherein the oligomer is labeled with at least one detectable moiety.
5. The PNA oligomer of claim 4 , wherein the detectable moiety or moieties are each independently selected from the group consisting of: a dextran conjugate, a branched nucleic acid detection system, a chromophore, a fluorophore, a spin label, a radioisotope, an enzyme, a hapten, an acridinium ester and a chemiluminescent compound.
6. The PNA oligomer of claim 5 , wherein the enzyme is selected from the group consisting of alkaline phosphatase, soybean peroxidase, horseradish peroxidase, ribonuclease and protease.
7. The PNA oligomer of claim 5 , wherein the hapten is selected from the group consisting of fluorescein, biotin, 2,4-dinitrophenyl and digoxigenin.
8. The PNA oligomer of claim 1 , wherein the oligomer is labeled with at least two independently detectable moieties.
9. The PNA oligomer of claim 8 , wherein the two or more independently detectable moieties are independently detectable fluorophores.
10. The PNA oligomer of claim 1 , wherein the oligomer comprises a non-fluorescent quencher moiety.
11. The PNA oligomer of claim 1 , wherein the oligomer comprises an energy transfer set of labels.
12. The PNA oligomer of claim 1 , wherein the oligomer is support bound.
13. A PNA oligomer that:
a) comprises one or more non-natural nucleobases comprising a sequence that is at least ninety percent homologous to a nucleobase sequence, or its complement, selected from the group consisting of: CCA-SSG-GTA-TCD-DTC (Seq. ID No. 1) and TTC-AAA-GGC-TCC-CGC (Seq. ID No. 2);
b) does not substantially intra or inter molecularly self-hybridize; and
c) sequence-specifically hybridizes to a region of the nucleic acid of Bacillus anthracis.
14. The PNA oligomer of claim 13 , wherein the non-natural nucleobases are each independently selected from the group consisting of 2,6-diaminopurine, 2-thiouracil and 2-thiothymine.
15. The PNA oligomer of claim 13 , wherein the oligomer is unlabeled.
16. The PNA oligomer of claim 13 , wherein the oligomer is labeled with at least one detectable moiety.
17. The PNA oligomer of claim 16 , wherein the detectable moiety or moieties are each independently selected from the group consisting of: a dextran conjugate, a branched nucleic acid detection system, a chromophore, a fluorophore, a spin label, a radioisotope, an enzyme, a hapten, an acridinium ester and a chemiluminescent compound.
18. The PNA oligomer of claim 17 , wherein the enzyme is selected from the group consisting of alkaline phosphatase, soybean peroxidase, horseradish peroxidase, ribonuclease and protease.
19. The PNA oligomer of claim 17 , wherein the hapten is selected from the group consisting of fluorescein, biotin, 2,4-dinitrophenyl and digoxigenin.
20. The PNA oligomer of claim 13 , wherein the oligomer is labeled with at least two independently detectable moieties.
21. The PNA oligomer of claim 20 , wherein the two or more independently detectable moieties are independently detectable fluorophores.
22. The PNA oligomer of claim 13 , wherein the oligomer comprises a non-fluorescent quencher moiety.
23. The PNA oligomer of claim 13 , wherein the oligomer comprises an energy transfer set of labels.
24. The PNA oligomer of claim 13 , wherein the oligomer is support bound.
25. An oligomer set comprising two or more oligomers, at least one of which is a PNA oligomer comprising a probing nucleobase sequence that is at least ninety percent homologous to a nucleobase sequence, or its complement, selected from the group consisting of: CCA-SSG-GTA-TCD-DTC (Seq. ID No. 1) and TTC-AAA-GGC-TCC-CGC (Seq. ID No. 2).
26. A method comprising: a) contacting a sample, under suitable hybridization conditions, with at least one PNA oligomer comprising a probing nucleobase sequence that is at least ninety percent homologous to a nucleobase sequence, or its complement, selected from the group consisting of: CCA-SSG-GTA-TCD-DTC (Seq. ID No. 1) and TTC-AAA-GGC-TCC-CGC (Seq. ID No. 2); and b) detecting, identify and/or quantitating hybridization of the probing nucleobase sequence of a PNA oligomer to a target sequence within the nucleic acid of Bacillus anthracis and correlating the result with the presence, absence and/or quantity of Bacillus anthracis in the sample.
27. A kit comprising:
a) one or more PNA oligomers comprising a probing nucleobase sequence that is at least ninety percent homologous to a nucleobase sequence, or its complement, selected from the group consisting of: CCA-SSG-GTA-TCD-DTC (Seq. ID No. 1), TTC-AAA-GGC-TCC-CGC (Seq. ID No. 2) and TTC-AAT-GGC-TCC-CGC (Seq. ID No. 3); and b) other reagents, instructions or compositions useful in performing an assay suitable for determining the presence, absence and/or quantity of the nucleic acid of Bacillus anthracis in a sample.