Compositions, methods and kits to detect herpes simplex virus nucleic acids

Patent metadata

Field	Value
Publication number	US-10073094-B2
Application number	US-201715487240-A
Country	US
Kind code	B2
Filing date	Apr 13, 2017
Priority date	Apr 24, 2012
Publication date	Sep 11, 2018
Grant date	Sep 11, 2018

Abstract

The disclosed invention is related to methods, compositions, kits and isolated nucleic acid sequences for targeting Herpes Simplex Virus (HSV) nucleic acid (e.g., HSV-1 and/or HSV-2 nucleic acid). Compositions include amplification oligomers, detection probe oligomers and/or target capture oligomers. Kits and methods comprise at least one of these oligomers.

First claim

The invention claimed is:

1. A method for determining the presence or absence of Herpes Simplex Virus 2 (HSV-2) in a sample, said method comprising:
(1) contacting a sample, said sample suspected of containing HSV-2, with at least two oligomers for amplifying a target region of an HSV-2 target nucleic acid, wherein the at least two amplification oligomers comprise
(a) a first amplification oligomer comprising a first target-hybridizing sequence that is from about 15 to about 27 contiguous nucleotides (i) contained in the sequence of SEQ ID NO:49 and that includes at least the sequence of SEQ ID NO:48 or (ii) contained in the sequence of SEQ ID NO:43 and that includes at least the sequence of SEQ ID NO:42; and
(b) a second amplification oligomer comprising a second target-hybridizing sequence that is from about 15 to about 27 contiguous nucleotides (i) contained in the sequence of SEQ ID NO:51 and that includes at least the sequence of SEQ ID NO:50 or (ii) contained in the sequence of SEQ ID NO:45 and that includes at least the sequence of SEQ ID NO:44;

(2) performing an in vitro nucleic acid amplification reaction, wherein any HSV-2 target nucleic acid present in said sample is used as a template for generating an amplification product; and
(3) detecting the presence or absence of the amplification product, thereby indicating the presence or absence of HSV-2 in said sample.

2. The method of claim 1 , wherein
the first target hybridizing sequence is contained in the sequence of SEQ ID NO:49 and includes at least the sequence of SEQ ID NO:48; and
the second target hybridizing sequence is contained in the sequence of SEQ ID NO:51 and includes at least the sequence of SEQ ID NO:50.

3. The method of claim 2 , wherein the first target-hybridizing sequence has the sequence of SEQ ID NO:24 and/or the second target hybridizing sequence has the sequence of SEQ ID NO: 25.

4. The method of claim 2 , wherein
the first target hybridizing sequence is contained in the sequence of SEQ ID NO:43 and includes at least the sequence of SEQ ID NO: 42; and
the second target hybridizing sequence is contained in the sequence of SEQ ID NO:45 and includes at least the sequence of SEQ ID NO:44.

5. The method of claim 4 , wherein the first target-hybridizing sequence has the sequence of SEQ ID NO:14.

6. The method of claim 4 , wherein the second target hybridizing sequence has the sequence of SEQ ID NO:15.

7. The method of claim 1 , wherein the first and second target-hybridizing sequences respectively have the nucleotide sequences of
(i) SEQ ID NO:24 and SEQ ID NO:25; or
(ii) SEQ ID NO:14 and SEQ ID NO:15.

8. The method of claim 1 , wherein the first amplification oligomer is a promoter primer or promoter provider further comprising a promoter sequence located 5′ to the first target-hybridizing sequence.

9. The method of claim 8 , wherein the promoter sequence is a T7 promoter sequence, optionally wherein the promoter sequence has the sequence of SEQ ID NO:54.

10. The method of claim 9 , wherein the first amplification oligomer has a sequence selected from the group consisting of SEQ ID NO:23 and SEQ ID NO:13.

11. The method of claim 1 , further comprising purifying the HSV-2 target nucleic acid from other components in the sample before step (1).

12. The method of claim 11 , wherein the purifying step comprises contacting the sample with at least one capture probe oligomer comprising (i) a target-hybridizing sequence covalently attached to a sequence or moiety that binds to an immobilized probe, wherein said target-hybridizing sequence comprises SEQ ID NO:4, SEQ ID NO:1, SEQ ID NO:70, or SEQ ID NO:72; or (ii) a sequence comprising SEQ ID NO:3, SEQ ID NO:17, SEQ ID NO:69, or SEQ ID NO:71; or (iii) a target-hybridizing sequence covalently attached to a sequence or moiety that binds to an immobilized probe, wherein said target-hybridizing sequence is from about 15 to about 30 contiguous nucleotides contained in the sequence of SEQ ID NO:76 and includes at least the sequence of SEQ ID NO:75; or (iv) a target-hybridizing sequence the is contained in the sequence of SEQ ID NO:74; or (v) a target-hybridizing sequence that is contained in the sequence of SEQ ID NO:74 and includes at least the sequence of SEQ ID NO:75.

13. The method of claim 1 , wherein the detecting step (3) comprises contacting said in vitro nucleic acid amplification reaction with a detection probe oligomer configured to specifically hybridize to the amplification product under conditions whereby the presence or absence of the amplification product is determined, thereby indicating the presence or absence of HSV-2 in said sample.

14. The method of claim 13 , wherein the detection probe oligomer comprises (i) a target-hybridizing sequence that is from about 14 to about 25 nucleotides in length and is configured to specifically hybridize to a target sequence contained within SEQ ID NO:2 from about nucleotide position 608 to about nucleotide position 632; or (ii) a target-hybridizing sequence that is contained in the sequence of SEQ ID NO:53 and includes at least the sequence of SEQ ID NO:52; or (iii) a target-hybridizing sequence consisting of SEQ ID NO:27; or (iv) a target-hybridizing sequence that is from about 14 to about 30 nucleotides in length and is configured to specifically hybridize to a target sequence contained within SEQ ID NO:2 from about nucleotide position 549 to about nucleotide position 578; or (v) a target-hybridizing sequence that is contained in the sequence of SEQ ID NO:47 and includes at least the sequence of SEQ ID NO:46; or (vi) a target-hybridizing sequence consisting of SEQ ID NO:16.

15. The method of claim 14 , wherein the detection probe further comprises a label, optionally wherein the label is a chemiluminescent label or a fluorescent label, further optionally wherein the label is a fluorescent label and the detection probe further comprises a quencher.

16. The method of claim 14 , wherein the detection probe further comprises a non-target-hybridizing sequence.

17. A combination of at least two oligomers for determining the presence or absence of Herpes Simplex Virus 2 (HSV-2) in a sample, said oligomer combination comprising:
first and second amplification oligomers for amplifying a target region of an HSV-2 target nucleic acid, wherein
(a) the first amplification oligomer comprises a first target-hybridizing sequence that is from about 15 to about 27 contiguous nucleotides (i) contained in the sequence of SEQ ID NO:49 and that includes at least the sequence of SEQ ID NO:48 or (ii) contained in the sequence of SEQ ID NO:43 and that includes at least the sequence of SEQ ID NO:42; and
(b) the second amplification oligomer comprises a second target-hybridizing sequence that is from about 15 to about 27 contiguous nucleotides (i) contained in the sequence of SEQ ID NO:51 and that includes at least the sequence of SEQ ID NO:50 or (ii) contained in the sequence of SEQ ID NO:45 and that includes at least the sequence of SEQ ID NO:44.

18. A reaction mixture comprising the combination of at least two oligomers as in claim 17 .

19. The combination of claim 17 , further comprising a detection probe oligomer configured to specifically hybridize to an amplification product produced by the first and second amplification oligomers in the presence of the under conditions whereby the presence or absence of the HSV-2 target nucleic acid, wherein the detection probe oligomer comprises a detectable label.

20. The combination of claim 17 , wherein the first amplifica

Assignees

• Gen Probe Inc

Inventors

• Getman Damon K
• Aiyer Aparna

Classifications

• A61K48/00

  Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• G01N33/569 Primary

  for microorganisms, e.g. protozoa, bacteria, viruses · CPC title

• C07K16/08

  against material from viruses · CPC title

• C12Q1/705 Primary

  for herpetoviridae, e.g. herpes simplex, varicella zoster · CPC title