Compositions, kits and methods for detection of campylobacter nucleic acid
The disclosed invention is related to compositions, kits and methods comprising one or more oligomers targeting 16S rRNA target nucleic acid from Campylobacter species jejuni, coli and/or lari . Compositions include amplification oligomers, detection probe oligomers and/or target capture oligomers. Kits and methods comprise at least one of these oligomers.
1. A composition for use in a Campylobacter target nucleic acid amplification assay, the composition comprising at least two amplification oligomers capable of stably hybridizing to a C. jejuni target nucleic acid and a C. coli , a C. lari or a C. coli and a C. lari target nucleic acid and amplifying a Campylobacter target nucleic acid in an amplification reaction, wherein a first of said amplification oligomers comprises a contiguous target hybridizing sequence 15 to 45 nucleotides in length and configured to hybridize to a sequence in a region of a Campylobacter 16S rRNA gene corresponding to nucleotides 108 to 150 of SEQ ID NO:91 and wherein a second of said amplification oligomers comprises a contiguous target hybridizing sequence 15 to 45 nucleotides in length and configured to hybridize to a sequence in a region of the a Campylobacter 16S rRNA gene corresponding to nucleotides 170 to 226 of SEQ ID NO:91, and wherein said first of said amplification oligomers is a promoter based amplification oligomer further comprising a 5′ promoter sequence.
2. The composition of claim 1 , wherein said second of said amplification oligomers comprises a target hybridizing sequence configured to hybridize to a sequence in a region of a Campylobacter 16S rRNA gene selected from the group consisting of: a region corresponding to nucleotides 170 to 212 of SEQ ID NO:91; a region corresponding to nucleotides 184 to 212 of SEQ ID NO:91; and a region corresponding to nucleotides 170 to 205 of SEQ ID NO:91.
3. The composition of claim 2 , wherein said first of said amplification oligomers comprises a target hybridizing sequence that consists essentially of SEQ ID NO:27.
4. The composition of claim 2 , wherein said first of said amplification oligomers comprises a sequence consisting of SEQ ID NO:26.
5. The composition of claim 2 , wherein said first of said amplification oligomers comprises a target hybridizing sequence selected from the group consisting of SEQ ID NOS:19, 21, 23, 25, 27, 29, 31 and 33.
6. The composition of claim 2 , wherein said first of said amplification oligomers comprises a sequence consisting of SEQ ID NO:32.
7. The composition of claim 1 , wherein said first of said amplification oligomers comprises a sequence that is selected from the group consisting of SEQ ID NOS:18, 20, 22, 24, 26, 28, 30, 32 and 84-88.
8. The composition of claim 1 , wherein said second of said amplification oligomers contains a target hybridizing sequence consisting of a sequence selected from the group consisting of SEQ ID NOS:1 through 14.
9. The composition of claim 8 , wherein said first of said amplification oligomers consists essentially of a sequence selected from the group consisting of SEQ ID NO:26 and SEQ ID NO:32.
10. The composition of claim 1 , wherein said second of said amplification oligomers consists essentially of a sequence of SEQ ID NO:7.
11. The composition of claim 1 , wherein said 5′ promoter sequence is SEQ ID NO:75.
12. The composition of claim 1 , further comprising a blocker oligomer wherein said blocker oligomer contains one or more blocking groups located at or near the 3′ end.
13. The composition of claim 1 , further comprising a detection oligomer comprising a sequence configured to hybridize to a portion of an amplification product generated from a target nucleic acid by said at least two amplification oligomers.
14. The composition of claim 13 , wherein said detection oligomer sequence is selected from the group consisting of SEQ ID NOS:63 through 68.
15. The composition of claim 13 , wherein said detection oligomer sequence consists essentially of SEQ ID NO:67 or consists essentially of SEQ ID NO:68.
16. The composition of claim 1 , wherein said first of said amplification oligomers comprises a sequence consisting essentially of a sequence selected from the group consisting of SEQ ID NO:26, SEQ ID NO:32, and SEQ ID NOS:84-88; and wherein said second amplification oligomer comprises a sequence consisting essentially of SEQ ID NO:7.
17. The composition of claim 16 , further comprising a detection oligomer comprising a sequence selected from the group consisting of SEQ ID NO:67 and SEQ ID NO:68.
18. The composition of claim 17 , wherein said first of said amplification oligomers comprises a sequence consisting essentially of a sequence selected from the group consisting of SEQ ID NO:26 and SEQ ID NO:32, and said composition further comprises a blocker oligomer comprises a sequence consisting of SEQ ID NO:50.
19. A reaction mixture containing the composition of claim 1 .