Kits for detecting group B Streptococci

• US 8,097,409 B2
• Filed: 02/23/2009
• Issued: 01/17/2012
• Est. Priority Date: 02/07/2005
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Pollner, Reinhold B., Kamantigue, Edgar J.
• Primary Examiner(s): OGUNBIYI, OLUWATOSIN A
• Application Number:
  US12/391,123
• Publication Number:
  US 20090176241A1
• Time in Patent Office: 1,058 Days
• Field of Search: None
• 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 for amplifying and detecting a Streptococcus agalactiae target nucleic acid sequence, said kit comprising:

a first primer, the base sequence of which consists of a 3′

terminal target-complementary sequence and a first primer upstream sequence comprising a promoter sequence for an RNA polymerase, said 3′

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

2 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs;

a second primer, the base sequence of which consists of a 3′

terminal target-complementary sequence contained within and comprising at least 20 contiguous bases of SEQ ID NO;

1 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs; and

a probe, the base sequence of which consists of a target-complementary sequence and, optionally, one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement, said target-complementary sequence of said probe being contained within and comprising at least 16 contiguous bases of SEQ ID NO;

3 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.

2. The kit of claim 1, 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, SEQ ID NO;

10, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.

3. The kit of claim 2, 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, SEQ ID NO;

6, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.

4. The kit of claim 1, 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, SEQ ID NO;

6, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.

5. The kit of claim 1, 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, SEQ ID NO;

17, and the full complements thereof, allowing for the presence of DNA base equivalents and nucleotide analogs.

6. The kit of claim 1, wherein said probe comprises said optional one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement.

7. The kit of claim 6, 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, SEQ ID NO;

17, and the full complements thereof, allowing for the presence of DNA base equivalents and nucleotide analogs.

8. The kit of claim 6, wherein said probe further comprises a fluorophore moiety and a quencher moiety.

9. A kit for amplifying and detecting a Streptococcus agalactiae target nucleic acid sequence, said kit comprising:

a first primer, the base sequence of which consists of a 3′

terminal target-complementary sequence and a first primer upstream sequence comprising a promoter sequence for an RNA polymerase, said 3′

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

8, SEQ ID NO;

9, SEQ ID NO;

10, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs;

a second primer, the base sequence of which consists of a 3′

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

4, SEQ ID NO;

5, SEQ ID NO;

6, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs; and

a probe, the base sequence of which consists of a target-complementary sequence and, optionally, one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement, said target-complementary sequence of said probe being selected from the group consisting of SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

17, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.

10. The kit of claim 9, wherein said probe comprises said optional one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement.

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

12. The kit of claim 9, wherein:

said 3′

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

10 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs;

said 3′

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

4 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs; and

said target-complementary sequence of said probe is SEQ ID NO;

16 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.

13. The kit of claim 12, wherein said probe comprises said optional one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement.

14. The kit of claim 13, wherein said probe further comprises a fluorophore moiety and a quencher moiety.