Kits for detecting group B
View Patent ↗Compositions, methods and kits for detecting Group B streptococci. Particularly described are oligonucleotides that are useful as amplification primers and hybridization probes for detecting very low levels of Group B streptococci nucleic acids.
1. A kit for amplifying and detecting a Streptococcus agalactiae target nucleic acid sequence, said kit comprising:
a first primer, the base sequence of which consists of a 3′ terminal target-complementary sequence and a first primer upstream sequence comprising a promoter sequence for an RNA polymerase, said 3′ terminal target-complementary sequence of said first primer being contained within and comprising at least 25 contiguous bases of SEQ ID NO:2 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs;
a second primer, the base sequence of which consists of a 3′ terminal target-complementary sequence contained within and comprising at least 20 contiguous bases of SEQ ID NO:1 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs; and
a probe, the base sequence of which consists of a target-complementary sequence and, optionally, one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement, said target-complementary sequence of said probe being contained within and comprising at least 16 contiguous bases of SEQ ID NO:3 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.
2. The kit of claim 1 , wherein said 3′ terminal target-complementary sequence of said first primer is selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.
3. The kit of claim 2 , wherein said 3′ terminal target-complementary sequence of said second primer is selected from the group consisting of SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.
4. The kit of claim 1 , wherein said 3′ terminal target-complementary sequence of said second primer is selected from the group consisting of SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.
5. The kit of claim 1 , wherein said target-complementary sequence of said probe is selected from the group consisting of SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, and the full complements thereof, allowing for the presence of DNA base equivalents and nucleotide analogs.
6. The kit of claim 1 , wherein said probe comprises said optional one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement.
7. The kit of claim 6 , wherein said target-complementary sequence of said probe is selected from the group consisting of SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, and the full complements thereof, allowing for the presence of DNA base equivalents and nucleotide analogs.
8. The kit of claim 6 , wherein said probe further comprises a fluorophore moiety and a quencher moiety.
9. A kit for amplifying and detecting a Streptococcus agalactiae target nucleic acid sequence, said kit comprising:
a first primer, the base sequence of which consists of a 3′ terminal target-complementary sequence and a first primer upstream sequence comprising a promoter sequence for an RNA polymerase, said 3′ terminal target-complementary sequence of said first primer being selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs;
a second primer, the base sequence of which consists of a 3′ terminal target-complementary sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs; and
a probe, the base sequence of which consists of a target-complementary sequence and, optionally, one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement, said target-complementary sequence of said probe being selected from the group consisting of SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, and the full complements thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.
10. The kit of claim 9 , wherein said probe comprises said optional one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement.
11. The kit of claim 10 , wherein said probe further comprises a fluorophore moiety and a quencher moiety.
12. The kit of claim 9 , wherein:
said 3′ terminal target-complementary sequence of said first primer is SEQ ID NO:10 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs;
said 3′ terminal target-complementary sequence of said second primer is SEQ ID NO:4 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs; and
said target-complementary sequence of said probe is SEQ ID NO:16 or the full complement thereof, allowing for the presence of RNA base equivalents and nucleotide analogs.
13. The kit of claim 12 , wherein said probe comprises said optional one or more base sequences that are non-complementary to a nucleic acid containing said target nucleic acid sequence or its complement.
14. The kit of claim 13 , wherein said probe further comprises a fluorophore moiety and a quencher moiety.