Highly conserved tuf genes and their use to generate probes and primers for detection of coagulase-negative
Provided herein are compositions and methods for the detection of Streptococcus agalacticae.
1. A method for specifically detecting the presence or absence in a sample of a tuf nucleic acid sequence belonging to a coagulase negative Staphylococcus comprising:
i) contacting the sample with an oligonucleotide probe or primer that hybridizes to a tuf nucleic acid sequence of each of SEQ ID NOs: 181, 182, 184-188, and 190-202 at nucleotides corresponding to nucleotides 313-679 of SEQ ID NO: 179, or complements thereof, but does not hybridize to a tuf nucleic acid sequence of any of SEQ ID NOs: 176-180 under stringent hybridization conditions of 55° C., 1.5 M NaCl and 10 mM EDTA;
ii) allowing said oligonucleotide to hybridize with said sample under conditions such that said oligonucleotide hybridizes to a tuf nucleic acid sequence of any of SEQ ID NOs: 181, 182, 184-188, and 190-202 at nucleotides corresponding to nucleotides 313-679 of SEQ ID NO: 179, or complements thereof, if present, but does not hybridize to a tuf nucleic acid sequence of any of SEQ ID NOs: 176-180 if present; and
iii) testing for hybridization of said oligonucleotide to a tuf nucleic acid sequence of SEQ ID NOs: 181, 182, 184-188, and 190-202 or complements thereof in said sample.
2. The method of claim 1 , wherein the nucleic acid sequence of said oligonucleotide comprises SEQ ID NO: 1175 or 1176 or a complement thereof.
3. The method of claim 1 , further comprising amplifying said tuf nucleic acid prior to hybridization of said oligonucleotide to said tuf nucleic acid.
4. The method of claim 3 , wherein said tuf nucleic acid is amplified using a first primer comprising the nucleic acid sequence of SEQ ID NO: 553 and a second primer comprising a nucleic acid sequence of SEQ ID NO: 575 or 707.
5. The method of claim 3 , wherein said oligonucleotide is a probe comprising the nucleic acid sequence of SEQ ID NO: 1175 or 1176.
6. The method of claim 5 , wherein said oligonucleotide comprises the nucleic acid sequence of SEQ ID NO: 1233.
7. The method of claim 4 , wherein said oligonucleotide is a probe comprising the nucleic acid sequence of SEQ ID NO: 1175 or 1176.
8. The method of claim 7 , wherein said oligonucleotide comprises the nucleic acid sequence of SEQ ID NO: 1233.
9. The method of claim 1 , wherein the nucleic acid sequence of said oligonucleotide consists of SEQ ID NO: 1175 or 1176 or a complement thereof.