Compositions and methods for detecting streptococcus agalactiae surface immunogenic protein genes
First Claim
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1. An oligonucleotide probe for detecting Streptococcus agalactia in a sample comprising a nucleotide sequence having at least 90% identity with a sequnce selected from the group consisting of:
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Abstract
This invention relates generally to Group B streptococci (GBS) Streptococcus agalactiae detection. More specifically, the present invention provides for novel probes for a specific and sensitive diagnostic test of GBS. These GBS-specific probes hybridize with the the surface immunogenic protein (sip) gene. Arrays comprising the probes immobilized on a support for hybridization analysis and methods for GBS detection using the probes are also provided.
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Citations
33 Claims
- 1. An oligonucleotide probe for detecting Streptococcus agalactia in a sample comprising a nucleotide sequence having at least 90% identity with a sequnce selected from the group consisting of:
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14. A method for detecting Streptococcus agalactia in a sample, comprising the steps of:
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a) providing an oligonucleotide probe comprising a nucleotide sequence, or a complementary strand thereof, having at least 90% identity with a sequence selected from the group consisting of;
5′
-ATCGACAATGGCAGCTTCGC-3′
,(SEQ ID NO;
6)5′
-CTGGTCAAACAACAGCTACTG-3′
,(SEQ ID NO;
7)5′
-AACAGGTATCACCAGCTCCT-3′
,(SEQ ID NO;
8)5′
-GAACAGGTATCACCAGCTCCTG-3′
,(SEQ ID NO;
9)5′
-GCAAGTGTTAAAGTAGTCACTC-3′
,(SEQ ID NO;
10)5′
-GGCAAGTGTTAAAGTAGTCACTCC-3′
,(SEQ ID NO;
11)3′
-TAGCTGTTACCGTCGAAGCG-5′
,(SEQ ID NO;
12)3′
-GACCAGTTTGTTGTCGATGAC-5′
,(SEQ ID NO;
13)3′
-TTGTCCATAGTGGTCGAGGT-5′
,(SEQ ID NO;
14)3′
-CTTGTCCATAGTCCTCGAGGAC-5′
,(SEQ ID NO;
15)3′
-CGTTCACAATTTCATCAGTGAG-5′
,(SEQ ID NO;
16)and 3′
-CCGTTCACAATTTCATCTGTGAGG-5′
;(SEQ ID NO;
17)b) contacting said probe with a sample containing or suspected of containing a Streptococcus agalactia target nucleotide sequence under conditions suitable for hybridization between said probe and said target nucleotide sequence; and
c) assessing hybridization between said probe and said target nucleotide sequence to detect said Streptococcus agalactia in said sample. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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27. A method for detecting Streptococcus agalactia in a sample comprising the steps of:
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a) providing an oligonucleotide probe comprising a nucleotide sequence that hybridizes with a target nucleotide sequence, wherein the target nucleotide sequence is all or part of the sip gene, or a complementary strand thereof, and wherein the probe has a G+C content from about 30 to 70%, a Tm value from about 40 to 80°
C., a length of at least 8 nucleotides and does not contain any hairpin secondary structure;
b) contacting said, probe with a sample containing or suspected of containing a Streptococcus agalactia target nucleotide sequence under conditions suitable for hybridization between said probe and said target nucleotide sequence; and
c) assessing hybridization between said probe and said target nucleotide sequence to detect said Streptococcus agalactia in said sample.
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28. A method for detecting Streptococcus agalactia in a sample comprising the steps of:
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a) providing an oligonucleotide probe comprising a nucleotide sequence that hybridizes under high stringency with a target comprising a nucleotide sequence selected from the group consisting of;
5′
-ATCGACAATGGCAGCTTCGC-3′
,(SEQ ID NO;
6)5′
-CTGGTCAAACAACAGCTACTG-3′
,(SEQ ID NO;
7)5′
-AACAGGTATCACCAGCTCCT-3′
,(SEQ ID NO;
8)5′
-GAACAGGTATCACCAGCTCCTG-3′
,(SEQ ID NO;
9)5′
-GCAAGTGTTAAAGTAGTCACTC-3′
,(SEQ ID NO;
10)5′
-GGCAAGTGTTAAAGTAGTCACTCC-3′
,(SEQ ID NO;
11)5′
-TAGCTGTTACCGTCGAAGCG-3′
,(SEQ ID NO;
12)3′
-TAGCTGTTACCGTCGAAGCG-5′
,(SEQ ID NO;
12)3′
-GACCAGTTTGTTGTCGATGAC-5′
,(SEQ ID NO;
13)3′
-TTGTCCATAGTGGTCGAGGT-5′
,(SEQ ID NO;
14)3′
-CTTGTCCATAGTCCTCGAGGAC-5′
,(SEQ ID NO;
15)3′
-CGTTCACAATTTCATCAGTGAG-5′
,(SEQ ID NO;
16)and 3′
-CCGTTCACAATTTCATCTGTGAGG-5′
;(SEQ ID NO;
17)b) contacting said probe with a sample containing or suspected of containing a Streptococcus agalactia target nucleotide sequence under conditions suitable for hybridization between said probe and said target nucleotide sequence; and
c) assessing hybridization between said probe and said target nucleotide sequence to detect said Streptococcus agalactia in said sample.
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- 29. An oligonucleotide probe for detecting Streptococcus agalactia in a sample comprising a nucleotide sequence that hybridizes with a target nucleotide sequence, or a complementary strand thereof, selected from the group consisting of:
Specification