Sequences and their use for detection and characterization of STEC bacteria
AssigneeE. I. Du Pont de Nemours and Company
InventorsStephen Varkey, Daniel R. DeMarco, Mark A. Jensen
Life of this patent
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Abstract
This invention relates to a rapid method for detection and characterization of STEC bacteria based on the presence of nucleic acid sequences, in particular, to a PCR-based method for detection, and to oligonucleotide molecules and reagents and kits useful therefore. This method is preferably employed to detect STEC bacteria in a food or water sample, such as a beef enrichment. The present invention further relates to isolated polynucleotides, replication compositions, kits, and reagent tablets for carrying out the method of the present invention.
Claims14
A method for detecting the presence of STEC bacteria in a sample, said sample comprising nucleic acids, said method comprising:
- (a) providing a reaction mixture comprising a first primer, a second primer, and a probe for amplification and detection of at least a portion of:
- (i) SEQ ID NO:
- 688;
- wherein each of said first primer for the amplification of SEQ ID NO:
- 688, second primer for the amplification of SEQ ID NO:
- 688, and probe for the detection of SEQ ID NO:
- 688 comprises a 5′ end and a 3′ end;
- wherein said first primer for the amplification of SEQ ID NO:
- 688 comprises at least 15 contiguous nucleotides of SEQ ID NO:
- 145 or a sequence complementary thereto;
- and wherein said probe for the detection of SEQ ID NO:
- 688 comprises at least 15 contiguous nucleotides of SEQ ID NO:
- 160 or a sequence complementary thereto;
- and
- (ii) one or more nucleic acid sequences selected from the group consisting of:
- (A) SEQ ID NO:
- 686;
- wherein each of said first primer for the amplification of SEQ ID NO:
- 686, second primer for the amplification of SEQ ID NO:
- 686, and probe for the detection of SEQ ID NO:
- 686 comprises a 5′ end and a 3′ end;
- and wherein said first primer for the amplification of SEQ ID NO:
- 686 and probe for the detection of SEQ ID NO:
- 686 are selected from the group consisting of:
- (I) a first primer comprising at least 15 contiguous nucleotides of SEQ ID NO:
- 1 or a sequence complementary thereto and a probe comprising at least 15 contiguous nucleotides of SEQ ID NO:
- 16 or a sequence complementary thereto;
- (II) a first primer comprising at least 15 contiguous nucleotides of SEQ ID NO:
- 1 or a sequence complementary thereto and a probe comprising at least 15 contiguous nucleotides of SEQ ID NO:
- 49 or a sequence complementary thereto;
- and
- (III) a first primer comprising SEQ ID NO:
- 55 or a sequence complementary thereto and probe comprising SEQ ID NO:
- 56 or a sequence complementary thereto;
- and
- (B) SEQ ID NO:
- 687;
- wherein each of said first primer for the amplification of SEQ ID NO:
- 687, second primer for the amplification of SEQ ID NO:
- 687, and probe for the detection of SEQ ID NO:
- 687 comprises a 5′ end and a 3′ end;
- and wherein said first primer for the amplification of SEQ ID NO:
- 687 and probe for the detection of SEQ ID NO:
- 687 are selected from the group consisting of:
- (I) a first primer comprising at least 15 contiguous nucleotides of SEQ ID NO:
- 74 or a sequence complementary thereto and a probe comprising at least 15 contiguous nucleotides of SEQ ID NO:
- 89 or a sequence complementary thereto;
- (II) a first primer comprising SEQ ID NO:
- 120 or a sequence complementary thereto and a probe comprising SEQ ID NO:
- 121 or 122 or a sequence complementary thereto;
- and
- (III) a first primer comprising SEQ ID NO:
- 125 or a sequence complementary thereto and probe comprising SEQ ID NO:
- 126 or a sequence complementary thereto;
- and
- (b) performing PCR amplification of said nucleic acids of said sample using the reaction mixture of step (a);
- and
- (c) detecting the amplification of step (b).
Dependencies are inferred from the claim text.