Novel serotype streptococcus mutans and utilization of the same
A polynucleotide encoding a polypeptide which lowers the glucose side chain amount in a serotype-specific polysaccharide antigen of Streptococcus mutans ; novel Streptococcus mutans strain having the above polynucleotide; an antibody specific to this Streptococcus mutans strain; and a method of detecting the above-described Streptococcus mutans strain occurring in a subject sample. According to this method, it is possible to examine the presence or absence of Streptococcus mutans of an untypable serotype in a subject sample to thereby specify a subject having a high risk of the onset of infective endocarditis.
1 . A polynucleotide that encodes a polypeptide that reduces the amount of glucose side chain of a polysaccharide antigen specific to Streptococcus mutans,
wherein the polynucleotide comprises:
a base sequence of any of SEQ ID NO: 1 through 4; or
a base sequence with the deletion, substitution, or addition of one or more bases in the base sequence of any of SEQ ID NO: 1 through 4.
2 . A polynucleotide that encodes a polypeptide that reduces the amount of glucose side chain of a polysaccharide antigen specific to Streptococcus mutans,
wherein the polynucleotide comprises:
a polynucleotide of a base sequence of any of SEQ ID NO: 1 through 4; or
a polynucleotide that hybridizes under stringent conditions with a polynucleotide having a complementary base sequence to the polynucleotide of the base sequence of any of SEQ ID NO: 1 through 4.
3 . An oligonucleotide that comprises a base sequence, or a complementary sequence thereof, with at least 12 contiguous bases of a base sequence of any of SEQ ID NO: 1 through 4.
4 . An oligonucleotide of claim 3 , which comprises a base sequence of any of SEQ ID NO: 8 through 10.
5 . A polypeptide that is encoded by a polynucleotide of claim 1 .
6 . A Streptococcus mutans strain with a reduced amount of glucose side chain in a polysaccharide antigen specific to Streptococcus mutans.
7 . A Streptococcus mutans strain of claim 6 , which comprises a polynucleotide comprising a base sequence of any of SEQ ID NO: 1 through 4; or a base sequence with the deletion, substitution, or addition of one or more bases in the base sequence of any of SEQ ID NO: 1 through 4.
8 . A Streptococcus mutans strain of claim 6 , which expresses a polypeptide encoded by a polynucleotide of a base sequence of any of SEQ ID NO: 1 through 4; or a polynucleotide that hybridizes under stringent conditions with a polynucleotide having a complementary base sequence to the polynucleotide of the base sequence of any of SEQ ID NO: 1 through 4.
9 . An antibody that specifically binds to a Streptococcus mutans strain of claim 6 .
10 . A method for detecting a Streptococcus mutans strain in a subject sample, comprising the steps of:
separating bacteria from the subject sample;
extracting genomic DNA or total RNA of the bacteria separated from the subject sample; and
carrying out a PCR reaction, using the genomic DNA or total RNA as a template, and using an oligonucleotide of claim 3 as a primer.
11 . A method for detecting a Streptococcus mutans strain in a subject sample set forth in claim 10 , wherein the tissue sample is obtained from blood, saliva, or plaque.
12 . A method for detecting a Streptococcus mutans strain in a subject sample set forth in claim 10 , wherein the primers are an oligonucleotide of a base sequence of SEQ ID NO: 8, and an oligonucleotide of a base sequence of SEQ ID NO: 9 or 10.
13 . A method for detecting a Streptococcus mutans strain in a subject sample, comprising the steps of:
separating bacteria from the subject sample;
extracting genomic DNA or total RNA of the bacteria separated from the subject sample; and
carrying out a hybridization reaction for the genomic DNA or total RNA, using an oligonucleotide of claim 3 as a probe.
14 . A method for detecting a Streptococcus mutans strain in a subject sample set forth in claim 13 , wherein the tissue sample is obtained from blood, saliva, or plaque.
15 . A method for detecting a Streptococcus mutans strain in a subject sample set forth in claim 13 , wherein the oligonucleotide has a base sequence of any of SEQ ID NO: 8 through 10.
16 . A method for detecting a Streptococcus mutans strain in a subject sample set forth in claim 10 , wherein the step of separating bacteria uses an antibody that specifically binds to a Streptococcus mutans strain with a reduced amount of glucose side chain in a polysaccharide antigen specific to Streptococcus mutans.
17 . A method for detecting a Streptococcus mutans strain in a subject sample, comprising the steps of:
separating bacteria from the subject sample;
incubating the separated bacteria with an antibody of claim 9 ; and
detecting bacteria that have bound to the antibody.
18 . A method for determining a serotype of a Streptococcus mutans strain in a subject sample, the method comprising using a method of claim 10 .
19 . A screening method of a Streptococcus mutans strain, the method comprising using a method of claim 10 .
20 . A Streptococcus mutans strain, which is obtained by a screening method of claim 19 .
21 . A kit for detecting a Streptococcus mutans strain, the kit comprising an oligonucleotide of claim 3 .
22 . A kit for detecting a Streptococcus mutans strain set forth in claim 21 , wherein the oligonucleotide comprises a base sequence of SEQ ID NO: 9.
23 . A kit for detecting a Streptococcus mutans strain set forth in claim 22 , the kit further comprising an oligonucleotide of a base sequence of SEQ ID NO: 8.
24 . A kit for detecting a Streptococcus mutans strain set forth in claim 23 , the kit further comprising an oligonucleotide of a base sequence of SEQ ID NO: 10.
25 . A kit for detecting a Streptococcus mutans strain set forth in claim 21 , which is used for a PCR reaction or a hybridization reaction.
26 . A kit for detecting a Streptococcus mutans strain, the kit comprising an antibody of claim 9 .
27 . A method for producing an antibody of claim 9 , comprising the step of injecting a Streptococcus mutans strain with a reduced amount of glucose side chain in a polysaccharide antigen specific to Streptococcus mutans , wherein said Streptococcus mutans strain is suspended in a buffer, intravenously into the auricular vein of rabbits repeatedly for 5 consecutive days.
28 . A method for producing an antibody set forth in claim 27 , further comprising the step of repeating immunization of the Streptococcus mutans strain, suspended in a phosphate-buffered saline, one week after the injecting step and for another 2 weeks, 5 times each week.
29 . A bacteria detecting tool, which comprises an oligonucleotide, fixed on a substrate, that include a base sequence with at least 12 contiguous bases of a base sequence of any of SEQ ID NO: 1 through 4.
30 . A bacteria detecting tool of claim 29 , wherein the oligonucleotide comprises a base sequence, or a complementary sequence thereof, with at least 12 continuous bases of a base sequence of any of SEQ ID NO: 1 through 4.