RAPID DETECTION OF THE "HIGH VIRULENT" ST-17 CLONE OF GROUP B STREPTOCOCCUS

• US 20140302498A1
• Filed: 04/16/2014
• Published: 10/09/2014
• 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: POYART, Claire, LAMY, Marie-Cecile, DRAMSI, Shaynoor, SAUVAGE, Elisabeth, GLASER, Philippe, TRIEU-CUOT, Patrick
• Granted Patent: US 9,551,039 B2
• Time in Patent Office: Days
• US Class Current: 435/6.11
• 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

1-55. -55. (canceled)

56. A method for the in vitro detection of a high-virulence group B streptococcus strain of S-17 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 nucleotide sequence of SEQ ID NO;

5, SEQ ID NO;

13, or SEQ ID NO;

15, or the complements thereof; and

detecting the presence or absence of a segment S10 or segment S11 nucleotide sequence in the nucleic acid of the biological sample.

57. The method of claim 56, further comprising hybridizing a positive control to the nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

5, SEQ ID NO;

13, or SEQ ID NO;

15, or the complements thereof.

58. The method of claim 56, further comprising hybridizing a negative control to the nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

5, SEQ ID NO;

13, or SEQ ID NO;

15, or the complements thereof.

59. The method of claim 57, further comprising hybridizing a negative control to the nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

5, SEQ ID NO;

13, or SEQ ID NO;

15, or the complements thereof.

60. The method of claim 56, comprising hybridizing the nucleic acid of the biological sample with a nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

5, or the complement thereof.

61. The method of claim 56, comprising hybridizing the nucleic acid of the biological sample with a nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

13, or the complement thereof.

62. The method of claim 56, comprising hybridizing the nucleic acid of the biological sample with a nucleic acid molecule that hybridizes to a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

15, or the complement thereof.

63. The method of claim 60, further comprising amplifying by the polymerase chain reaction (PCR).

64. The method of claim 61, further comprising amplifying by the polymerase chain reaction (PCR).

65. The method of claim 62, further comprising amplifying by the polymerase chain reaction (PCR).

66. The method of claim 60, 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.

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

providing a biological sample comprising nucleic acid;

amplifying a fragment of a segment S10 nucleotide sequence or a fragment of a segment S11 nucleotide sequence in the nucleic acid of the biological sample with primers that amplify a fragment of a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO;

5, SEQ ID NO;

13, or SEQ ID NO;

15; and

detecting the amplification product.

68. The method of claim 67, 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.

69. The method of claim 67, comprising amplifying a nucleic acid sequence within the segment S11 nucleic acid sequence, wherein the primers amplify a fragment of a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

13.

70. The method of claim 67, comprising amplifying a nucleic acid sequence within the segment S11 nucleic acid sequence, wherein the primers amplify a fragment of a nucleotide sequence consisting of the nucleotide sequence of SEQ ID NO:

15.

71. The method of claim 67, comprising amplifying by the polymerase chain reaction (PCR).

72. The method of claim 68, comprising amplifying by the polymerase chain reaction (PCR).

73. The method of claim 69, comprising amplifying by the polymerase chain reaction (PCR).

74. The method of claim 70, comprising amplifying by the polymerase chain reaction (PCR).

75. The method of claim 67, 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.

76. The method of claim 67, wherein both 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.

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

providing a biological sample comprising nucleic acid;

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

5;

subjecting the biological sample to an amplification reaction; and

detecting the presence or absence of a segment S10 sequence in the nucleic acid of the biological sample.

78. The method of claim 77, wherein the amplification reaction is a polymerase chain reaction (PCR).

79. The method of claim 77, wherein primer set has the nucleotide sequences of SEQ ID NO:

33 and SEQ ID NO;

34.