Rapid detection of the “high virulent” ST-17 clone of group B

• US 9,551,039 B2
• Filed: 04/16/2014
• Issued: 01/24/2017
• Est. Priority Date: 12/20/2005
• Status: Active Grant

• Current Assignee: Assistance Publique - Hopitaux De Paris, Centre National De La Recherche Scientifique, Institut Pasteur, Universite Paris CITE
• Original Assignee: Assistance Publique - Hopitaux De Paris, Centre National De La Recherche Scientifique, Institut Pasteur, Université Paris Descartes
• Inventors: Dramsi, Shaynoor, Sauvage, Elisabeth, Poyart, Claire, Lamy, Marie-Cecile, Glaser, Philippe, Trieu-Cuot, Patrick
• Primary Examiner(s): Dauner, Joseph G
• Application Number:
  US14/254,051
• Publication Number:
  US 20140302498A1
• Time in Patent Office: 1,014 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C07K 16/1275 from Streptococcus (G)
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: G01N 2333/315 from Streptococcus (G), e.g. Enterococci
• CPC Class Codes: G01N 33/56944 Streptococcus

Abstract

The present invention relates to polynucleotides enabling the rapid, simple and specific detection of Group B Streptococcus highly-virulent ST-17 clones.

The present invention also relates to the polypeptides encoded by said polynucleotides, as well as to antibodies directed or raised against said polypeptides.

The present invention also relates to kits and methods for the specific detection of Group B Streptococcus highly-virulent ST-17 clones, using the polynucleotides, the polypeptides or the antibodies according to the invention.

Claims

1. A method for the in vitro detection of a high-virulence group B streptococcus strain of the ST-17 clone comprising:

providing a biological sample comprising nucleic acid;

hybridizing the nucleic acid of the biological sample with a nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the S10 nucleotide sequence of SEQ ID NO;

5, the S11a nucleotide sequence of SEQ ID NO;

13, or the S11b nucleotide sequence of SEQ ID NO;

15, or the complements thereof; and

detecting the presence or absence of the ST-17 clone by detecting the presence or absence of the S10, S11a or S11b nucleotide sequence or the complement thereof in the nucleic acid of the biological sample.

2. The method of claim 1, further comprising hybridizing a positive control to the nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the S10 nucleotide sequence of SEQ ID NO:

5, the S11a nucleotide sequence of SEQ ID NO;

13 or the S11b nucleotide sequence of SEQ ID NO;

15, or the complements thereof.

3. The method of claim 2, further comprising hybridizing a negative control to the nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the S10 nucleotide sequence of SEQ ID NO:

5, the S11a nucleotide sequence of SEQ ID NO;

13 or the S11b nucleotide sequence of SEQ ID NO;

15, or the complements thereof.

4. The method of claim 1, further comprising hybridizing a negative control to the nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the S10 nucleotide sequence of SEQ ID NO:

5, the S11a nucleotide sequence of SEQ ID NO;

13 or the S11b nucleotide sequence of SEQ ID NO;

15, or the complements thereof.

5. The method of claim 1, comprising hybridizing the nucleic acid of the biological sample with a nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the S10 nucleotide sequence of SEQ ID NO:

5, or the complement thereof.

6. The method of claim 5, further comprising amplifying the nucleic acid of the biological sample using a polymerase chain reaction.

7. The method of claim 5, wherein the nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

5, or the complement thereof, is a primer consisting of the nucleotide sequence of SEQ ID NO;

33 or SEQ ID NO;

34.

8. The method of claim 1, comprising hybridizing the nucleic acid of the biological sample with a nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the S11a nucleotide sequence of SEQ ID NO:

13, or the complement thereof.

9. The method of claim 8, further comprising amplifying the nucleic acid of the biological sample using a polymerase chain reaction.

10. The method of claim 1, comprising hybridizing the nucleic acid of the biological sample with a nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the S11b nucleotide sequence of SEQ ID NO. 15, or the complement thereof.

11. The method of claim 10, further comprising amplifying the nucleic acid of the biological sample using a polymerase chain reaction.

12. A method for the in vitro detection of a high-virulence group B streptococcus strain of the ST-17 clone comprising:

providing a biological sample comprising nucleic acid;

amplifying a fragment of a segment S10 nucleotide sequence or a fragment of a segment S11a or segment S11b nucleotide sequence of the ST-17 clone in the nucleic acid of the biological sample with primers that amplify a fragment of the segment S10 nucleotide sequence of SEQ ID NO;

5, the segment S11a nucleotide sequence of SEQ ID NO;

13 or the segment S11b nucleotide sequence of SEQ ID NO;

15; and

detecting the ST-17 clone by detecting the S10, S11a or S11b amplification product.

13. The method of claim 12, comprising amplifying a nucleic acid sequence within the segment S10 nucleic acid sequence, wherein the primers amplify a fragment of a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

5.

14. The method of claim 13, wherein the amplifying comprises a polymerase chain reaction (PCR).

15. The method of claim 13, wherein at least one of the primers is selected from the group of primers consisting of the nucleotide sequences of SEQ ID NO:

33;

SEQ ID NO;

34;

SEQ ID NO;

28;

SEQ ID NO;

27; and

SEQ ID NO;

29.

16. The method of claim 13, wherein two of the primers are selected from the group primers consisting of the nucleotide sequences of SEQ ID NO:

33;

SEQ ID NO;

34;

SEQ ID NO;

28;

SEQ ID NO;

27; and

SEQ ID NO;

29.

17. The method of claim 16, wherein two of the primers are selected from the group primers consisting of the nucleotide sequences of SEQ ID NO:

33 and SEQ ID NO;

34.

18. The method of claim 12, comprising amplifying a nucleic acid sequence within the segment S11a nucleic acid sequence, wherein the primers amplify a fragment of a nucleotide sequence consisting of the segment S11a nucleotide sequence of SEQ ID NO:

13.

19. The method of claim 18, wherein the amplifying comprises a polymerase chain reaction (PCR).

20. The method of claim 12, comprising amplifying a nucleic acid sequence within the segment S11b nucleic acid sequence, wherein the primers amplify a fragment of a nucleotide sequence consisting of the segment S11b nucleotide sequence of SEQ ID NO:

15.

21. The method of claim 20, wherein the amplifying comprises a polymerase chain reaction (PCR).

22. The method of claim 12, wherein the amplifying comprises a polymerase chain reaction (PCR).

23. A method for the in vitro detection of a high-virulence group B streptococcus strain of the ST-17 clone comprising:

providing a biological sample comprising nucleic acid;

adding to the biological sample a primer set that amplifies a fragment of the S10 nucleotide sequence of SEQ ID NO;

5;

subjecting the biological sample to an amplification reaction; and

detecting the presence or absence of the ST-17 clone by detecting the presence or absence of an amplification product of the S10 nucleotide sequence in the nucleic acid of the biological sample.

24. The method of claim 23, wherein the amplification reaction is a polymerase chain reaction.

25. The method of claim 23, wherein primer set has the nucleotide sequences of SEQ ID NO:

33 and SEQ ID NO;

34.

26. A method for the in vitro detection of a high-virulence group B streptococcus strain of the ST-17 clone comprising:

providing a biological sample comprising nucleic acid;

adding to the biological sample a primer set that amplifies a fragment of the segment S10 nucleotide sequence of SEQ ID NO;

5, the segment S11a nucleotide sequence of SEQ ID NO;

13 or the segment S11b nucleotide sequence of SEQ ID NO;

15;

subjecting the biological sample to an amplification reaction; and

detecting the presence or absence of the ST-17 clone by detecting the presence or absence of an amplification product of the segment S10, segment S11a or segment S11b nucleotide sequence in the nucleic acid of the biological sample.