IP Library Granted Patent US 8,026,051
Granted Patent B2
US 8,026,051 · App. 10/150,045 · Granted Sep 27, 2011

PNA probes, probe sets, methods and kits pertaining to the detection of

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Quick Facts
Patent No.
US 8,026,051
App. No.
10/150,045
Granted
Sep 27, 2011
Kind
B2
Abstract

This invention is related to novel PNA probes, probe sets, methods and kits pertaining to the detection of one or more species of Candida yeast. Non-limiting examples of probing nucleobase sequences that can be used for the probes of this invention can be selected from the group consisting of: AGA-GAG-CAG-CAT-GCA (Seq. Id. No. 1), AGA-GAG-CAA-CAT-GCA (Seq. Id. No. 2), ACA-GCA-GAA-GCC-GTG (Seq. Id. No. 3), CAT-AAA-TGG-CTA-CCA-GA (Seq. Id. No. 4), CAT-AAA-TGG-CTA-CCC-AG (Seq. Id. No. 5), ACT-TGG-AGT-CGA-TAG (Seq. Id. No. 6), CCA-AGG-CTT-ATA-CTC-GC (Seq. Id. No. 7), CCC-CTG-AAT-CGG-GAT (Seq. Id. No. 8), GAC-GCC-AAA-GAC-GCC (Seq. Id. No. 9), ATC-GTC-AGA-GGC-TAT-AA (Seq. Id. No. 10), TAG-CCA-GAA-GAA-AGG (Seq. Id. No. 11), CAT-AAA-TGG-CTA-GCC-AG (Seq. Id. No. 12), CTC-CGA-TGT-GAC-TGC-G (Seq. Id. No. 13), TCC-CAG-ACT-GCT-CGG (Seq. Id. No. 14), TCC-AAG-AGG-TCG-AGA (Seq. Id. No. 15), GCC-AAG-CCA-CAA-GGA (Seq. Id. No. 16), GCC-GCC-AAG-CCA-CA (Seq. Id. No. 17), GGA-CTT-GGG-GTT-AG (Seq. Id. No. 18), CCG-GGT-GCA-TTC-CA (Seq. Id. No. 19), ATG-TAG-AAC-GGA-ACT-A (Seq. Id. No. 20), GAT-TCT-CGG-CCC-CAT-G (Seq. Id. No. 21), CTG-GTT-CGC-CAA-AAA-G (Seq. Id. No. 22) and AGT-ACG-CAT-CAG-AAA (Seq. Id. No. 23).

Claims (64)

1. A method comprising:

a) contacting a sample with a combination of PNA probes, wherein the combination of PNA probes comprises the probing nucleobase sequences consisting of SEQ ID NO: 1-23:

AGA-GAG-CAG-CAT-GCA,

(Seq. Id. No. 1)

AGA-GAG-CAA-CAT-GCA,

(Seq. Id. No. 2)

ACA-GCA-GAA-GCC-GTG,

(Seq. Id. No. 3)

CAT-AAA-TGG-CTA-CCA-GA,

(Seq. Id. No. 4)

CAT-AAA-TGG-CTA-CCC-AG,

(Seq. Id. No. 5)

ACT-TGG-AGT-CGA-TAG,

(Seq. Id. No. 6)

CCA-AGG-CTT-ATA-CTC-GC,

(Seq. Id. No. 7)

CCC-CTG-AAT-CGG-GAT,

(Seq. Id. No. 8)

GAC-GCC-AAA-GAC-GCC,

(Seq. Id. No. 9)

ATC-GTC-AGA-GGC-TAT-AA,

(Seq. Id. No. 10)

TAG-CCA-GAA-GAA-AGG,

(Seq. Id. No. 11)

CAT-AAA-TGG-CTA-GCC-AG,

(Seq. Id. No. 12)

CTC-CGA-TGT-GAC-TGC-G,

(Seq. Id. No. 13)

TCC-CAG-ACT-GCT-CGG,

(Seq. Id. No. 14)

TCC-AAG-AGG-TCG-AGA,

(Seq. Id. No. 15)

GCC-AAG-CCA-CAA-GGA,

(Seq. Id. No. 16)

GCC-GCC-AAG-CCA-CA,

(Seq. Id. No. 17)

GGA-CTT-GGG-GTT-AG,

(Seq. Id. No. 18)

CCG-GGT-GCA-TTC-CA,

(Seq. Id. No. 19)

ATG-TAG-AAC-GGA-ACT-A,

(Seq. Id. No. 20)

GAT-TCT-CGG-CCC-CAT-G,

(Seq. Id. No. 21)

CTG-GTT-CGC-CAA-AAA-G,

(Seq. Id. No. 22)

and

AGT-ACG-CAT-CAG-AAA,

(Seq. Id. No. 23)

or the complements thereof; and

b) detecting, identifying and/or quantifying hybridization of one or more of the PNA probing nucleobase sequences to the target sequence of one or more species of Candida yeast in the sample, under suitable hybridization conditions or suitable in-situ hybridization conditions, and correlating the result with the presence, absence and/or quantity of Candida yeast in the sample.

2. The method of claim 1 , wherein at least one of the PNA probes is unlabeled.

3. The method of claim 2 , wherein at least one PNA probe is support bound.

4. The method of claim 1 , wherein the PNA probes are all unlabeled.

5. The method of claim 1 , wherein at least one PNA probe is labeled with a detectable moiety.

6. The method of claim 5 , wherein the detectable moiety or moieties are 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.

7. The method of claim 5 , wherein in situ hybridization using a fluorophore or enzyme-linked probe is used to determine the presence, absence or number of organisms of Candida yeast in the sample.

8. The method of claim 5 , wherein one or more probes of the set are labeled with two or more independently detectable moieties.

9. The method of claim 8 , wherein the two or more independently detectable moieties are independently detectable fluorophores.

10. The method of claim 5 , wherein at least one PNA probe is self-indicating.

11. The method of claim 1 , wherein all probes of the set are labeled with one or more detectable moieties.

12. The method of claim 1 , wherein at least one PNA probe is labeled with at least two independently detectable moieties.

13. The method of claim 12 , wherein the two or more independently detectable moieties are independently detectable fluorophores.

14. The method of claim 1 , wherein two or more PNA probes are independently detectable.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 12, 2014
From: BOSTON PROBES, INC.
To: APPLIED BIOSYSTEMS, LLC
Reel/Frame 034627/0546 →
LIEN RELEASE Recorded Apr 8, 2013
From: BANK OF AMERICA, N.A.
To: BOSTON PROBES, INC.
Reel/Frame 030174/0230 →
SECURITY AGREEMENT Recorded Dec 5, 2008
From: BOSTON PROBES, INC.
To: BANK OF AMERICA, N.A., AS COLLATERAL AGENT
Reel/Frame 021924/0629 →