Compositions, kits and related methods for the detection and/or monitoring of
View Patent ↗Provided are compositions, kits, and methods for the identification of Salmonella . In certain aspects and embodiments, the compositions, kits, and methods may provide improvements in relation to specificity, sensitivity, and speed of detection.
1. A composition for use in a nucleic acid amplification assay to detect Salmonella in a sample, comprising a T7 provider oligonucleotide and a primer oligonucleotide that each target a sequence in a region of Salmonella nucleic acid,
wherein said T7 provider oligonucleotide and primer oligonucleotide used in said amplification assay target opposite strands of the Salmonella nucleic acid sequence to be amplified; and
wherein said T7 provider oligonucleotide targets a sequence in a region of Salmonella nucleic acid corresponding to bases from about 268-320 of E. coli 23S rRNA and/or said primer oligonucleotide targets a sequence in a region of Salmonella nucleic acid primer oligonucleotide targets a sequence in a region of Salmonella nucleic acid corresponding to bases 338-395 of E. coli 23S rRNA.
2. The composition of claim 1 , wherein said T7 provider oligonucleotide targets a sequence in a region of Salmonella nucleic acid corresponding to bases 268-302 or to bases 279-310 of E. coli 23S rRNA, and wherein said primer oligonucleotide targets a sequence in a region of Salmonella nucleic acid corresponding to bases 349-374 of E. coli 23S rRNA.
3. The composition of claim 1 or claim 2 , wherein said T7 provider oligonucleotide and said primer oligonucleotide comprises 15-35 nucleotides that are at least 90% complementary to the targeted Salmonella nucleic acid sequence.
4. The composition of claim 1 , wherein said T7 provider oligonucleotide is selected from the sequences of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, and 26.
5. The composition of claim 1 , wherein said primer oligonucleotide is selected from the sequences of SEQ ID NOs: 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, and 51.
6. The composition of claim 1 , wherein said T7 provider oligonucleotide is the sequence of SEQ ID NO: 17 or complement thereof, and said primer oligonucleotide is the sequence of SEQ ID NO: 50.
7. The composition of claim 1 , wherein said T7 provider oligonucleotide is the sequence of SEQ ID NO: 26 or complement thereof, and said primer oligonucleotide is the sequence of SEQ ID NO: 49.
8. The composition of claim 1 or claim 2 , further comprising a blocker oligonucleotide.
9. A kit for use in a Salmonella nucleic acid amplification assay comprising the composition according to claim 1 .
10. The kit of claim 9 , further comprising a detection oligonucleotide.
11. The kit of claim 10 , wherein said detection oligonucleotide is a torch oligonucleotide or molecular beacon.
12. A method for detecting Salmonella in a sample, said method comprising performing a nucleic acid amplification assay using the T7 provider oligonucleotide and the primer oligonucleotide composition of claim 1 .
13. The method of claim 12 , wherein detecting said amplified nucleic acid occurs in real-time.
14. The method of claim 12 or claim 13 , wherein Salmonella nucleic acid is specifically detected in the presence of at least one of: E. coli, Enterobacter, Citrobacter, Shigella, Pseudomonas and Listeria.
15. The method of claim 12 , wherein Salmonella nucleic acid is specifically detected in the presence of at least one of: E. coli, E. vulneris, E. hermannii, E. cloacae, E. aerogenes, E. hoshinae, P. mirabilis, C. brakii, P. fluorescens, S. flexneri, C. freundii, C. koseri/diversus, K. pneumoniae, S. marcescens, L. innocua, E. faecalis, C. jejuni, C. coli , and S. pneumoniae.