COMPOSITIONS AND METHODS FOR DETECTING GROUP B STREPTOCOCCI

• US 20090176241A1
• Filed: 02/23/2009
• Published: 07/09/2009
• Est. Priority Date: 02/07/2005
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Kamantigue, Edgar J., POLLNER, Reinhold B.
• Granted Patent: US 8,097,409 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6865 Promoter-based amplification, e.g. nucleic acid sequence amplification [NASBA], self-sustained sequence replication [3SR] or transcription-based amplification system [TAS]
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 2537/162 Helper probe

Abstract

Compositions, methods and kits for detecting Group B streptococci. Particularly described are oligonucleotides that are useful as amplification primers and hybridization probes for detecting very low levels of Group B streptococci nucleic acids.

Claims

1. A kit comprising:

a first primer that comprises a 3′

terminal target-complementary sequence and optionally a first primer upstream sequence that is not complementary to Streptococcus agalactiae nucleic acid sequence that is to be amplified, said 3′

terminal target-complementary sequence of said first primer comprising 25 contiguous bases contained within SEQ ID NO;

2, or the full complement thereof,

a second primer that comprises a 3′

terminal target-complementary sequence and optionally a second primer upstream sequence that is not complementary to Streptococcus agalactiae nucleic acid sequence that is to be amplified, said 3′

terminal target-complementary sequence of said second primer comprising 20 contiguous bases contained within SEQ ID NO;

1, or the full complement thereof; and

a probe that comprises a target-complementary sequence and optionally one or more base sequences that are not complementary to Streptococcus agalactiae nucleic acid sequence that is to be detected, said target-complementary sequence of said probe comprising 16 contiguous bases contained within SEQ ID NO;

3, or the full complement thereof.

2. The kit of claim 1, wherein said first primer and said second primer are each up to 58 bases in length.

3. The kit of claim 1, wherein said 3′

terminal target-complementary sequence of said first primer and said 3′

terminal target-complementary sequence of said second primer are each up to 31 bases in length.

4. The kit of claim 3, wherein said 3′

terminal target-complementary sequence of said second primer is up to 21 bases in length.

5. The kit of claim 1, wherein said first primer comprises said first primer upstream sequence.

6. The kit of claim 5, wherein said first primer upstream sequence comprises a promoter sequence for T7 RNA polymerase.

7. The kit of claim 3, wherein said 3′

terminal target-complementary sequence of said first primer is selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9 and SEQ ID NO;

10.

8. The kit of claim 4, wherein said 3′

terminal target-complementary sequence of said second primer is selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5 and SEQ ID NO;

6.

9. The kit of claim 1, wherein said probe is up to 30 bases in length.

10. The kit of claim 1, wherein said probe comprises said optional one or more base sequences that are not complementary to Streptococcus agalactiae nucleic acid sequence that is to be detected.

11. The kit of claim 10, wherein said probe further comprises a fluorophore moiety and a quencher moiety.

12. The kit of claim 11, wherein said target-complementary sequence of said probe is selected from the group consisting of SEQ ID NO:

14, SEQ ID NO;

15, SEQ ID NO;

16 and SEQ ID NO;

17.

13. A kit comprising:

a first primer that comprises a 3′

terminal target-complementary sequence and optionally a first primer upstream sequence that is not complementary to Streptococcus agalactiae nucleic acid sequence that is to be amplified, said 3′

terminal target-complementary sequence of said first primer selected from the group consisting of SEQ ID NO;

8, SEQ ID NO;

9 and SEQ ID NO;

10;

a second primer that comprises a 3′

terminal target-complementary sequence and optionally a second primer upstream sequence that is not complementary to Streptococcus agalactiae nucleic acid sequence that is to be amplified, said 3′

terminal target-complementary sequence of said second primer selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5 and SEQ ID NO;

6; and

a probe that comprises a target-complementary sequence and optionally one or more base sequences that are not complementary to Streptococcus agalactiae nucleic acid sequence that is to be detected, said target-complementary sequence of said probe selected from the group consisting of SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16 and SEQ ID NO;

17.

15. The kit of claim 13 or 14, wherein said first primer comprises said first primer upstream sequence.

16. The kit of claim 15, wherein said first primer upstream sequence comprises a promoter sequence for T7 RNA polymerase.

17. The kit of claim 13 or 14, wherein said probe comprises said optional one or more base sequences that are not complementary to Streptococcus agalactiae nucleic acid sequence that is to be detected.

18. The kit of claim 17, wherein said probe further comprises a fluorophore moiety and a quencher moiety.

14. A kit comprising:

a first primer that comprises a 3′

terminal target-complementary sequence and optionally a first primer upstream sequence that is not complementary to Streptococcus agalactiae nucleic acid sequence that is to be amplified, said 3′

terminal target-complementary sequence of said first primer consisting of SEQ ID NO;

10;

a second primer that comprises a 3′

terminal target-complementary sequence and optionally a second primer upstream sequence that is not complementary to Streptococcus agalactiae nucleic acid sequence that is to be amplified, said 3′

terminal target-complementary sequence of said second primer consisting of SEQ ID NO;

4; and

a probe that comprises a target-complementary sequence and optionally one or more base sequences that are not complementary to Streptococcus agalactiae nucleic acid sequence that is to be detected, said target-complementary sequence of said probe consisting of SEQ ID NO;

15.