Sequences and their use for detection and characterization of STEC bacteria
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.
1. An isolated polynucleotide comprising a primer-probe complex, wherein said primer probe complex comprises:
(A) a primer region comprising a nucleic acid sequence comprising at least 15 contiguous nucleotides of SEQ ID NO: 1 and a probe region comprising at least 15 contiguous nucleotides of SEQ ID NO: 16;
(B) a primer region comprising SEQ ID NO: 48 and a probe region comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 49-52;
(C) a primer region comprising SEQ ID NO: 55, and a probe region comprising SEQ ID NO: 56;
(D) a primer region comprising at least 15 contiguous nucleotides of SEQ ID NO: 74 and a probe region comprising at least 15 contiguous nucleotides of SEQ ID NO: 89;
(E) a primer region comprising SEQ ID NO: 120 and a probe region comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 121-122;
(F) a primer region comprising SEQ ID NO: 125 and a probe region comprising SEQ ID NO: 126; or
(G) a primer region comprising at least 15 contiguous nucleotides of SEQ ID NO: 145 and a probe region comprising at least 15 contiguous nucleotides of SEQ ID NO: 160;
wherein said probe region and said primer region each have a 5′ and 3′ terminus,
wherein said 3′ terminus of said probe region is attached to said 5′ terminus of said primer region via a linker moiety, and wherein said primer-probe complex further comprises a detectable label.
2. A kit for detection of STEC bacteria in a sample, comprising an isolated polynucleotide of claim 1 .
3. A reagent tablet comprising an isolated polynucleotide of claim 1 .