Nucleic acid amplification and detection of mycobacterium species
First Claim
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1. A method of detecting Mycobacterium species present in a biological sample, comprising the steps of:
- providing a biological sample containing nucleic acid from at least one Mycobacterium species comprising a Mycobacterium 16S ribosomal RNA (rRNA) or DNA encoding a Mycobacterium 16S rRNA;
amplifying the Mycobacterium 16S rRNA or Mycobacterium DNA encoding the Mycobacterium 16S rRNA in an in vitro nucleic acid amplification mixture comprising at least one polymerase activity, and a combination of at least one first oligonucleotide and at least one second oligonucleotide, wherein the first oligonucleotide consists a target-specific sequence that consists of SEQ ID NO;
5 joined to a 5′
promoter sequence or SEQ ID NO;
11, and the second oligonucleotide consists of SEQ ID NO;
16 or SEQ ID NO;
38 to produce amplified Mycobacterium nucleic acid; and
detecting the amplified Mycobacterium nucleic acid by detecting a label associated with the amplified Mycobacterium nucleic acid.
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Abstract
Oligonucleotides used to prime in vitro nucleic acid amplification of 16S rRNA sequences or DNA encoding 16S rRNA sequences for many species within the genus Mycobacterium are disclosed. Kits including such oligonucleotides are disclosed. Methods of detecting Mycobacterium species using the oligonucleotides in in vitro nucleic acid amplification are disclosed.
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Citations
20 Claims
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1. A method of detecting Mycobacterium species present in a biological sample, comprising the steps of:
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providing a biological sample containing nucleic acid from at least one Mycobacterium species comprising a Mycobacterium 16S ribosomal RNA (rRNA) or DNA encoding a Mycobacterium 16S rRNA; amplifying the Mycobacterium 16S rRNA or Mycobacterium DNA encoding the Mycobacterium 16S rRNA in an in vitro nucleic acid amplification mixture comprising at least one polymerase activity, and a combination of at least one first oligonucleotide and at least one second oligonucleotide, wherein the first oligonucleotide consists a target-specific sequence that consists of SEQ ID NO;
5 joined to a 5′
promoter sequence or SEQ ID NO;
11, and the second oligonucleotide consists of SEQ ID NO;
16 or SEQ ID NO;
38 to produce amplified Mycobacterium nucleic acid; anddetecting the amplified Mycobacterium nucleic acid by detecting a label associated with the amplified Mycobacterium nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A composition for amplifying in an in vitro amplification reaction a Mycobacterium 16S rRNA sequence or a DNA encoding 16S rRNA, comprising a combination of at least one first oligonucleotide and at least one second oligonucleotide, wherein the first oligonucleotide consists of a target-specific sequence that consists of SEQ ID NO:
- 5 joined to a 5′
promoter sequence or SEQ ID NO;
11, and wherein the second oligonucleotide consists of SEQ ID NO;
16 or SEQ ID NO;
38. - View Dependent Claims (14, 15, 16)
- 5 joined to a 5′
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17. A kit containing at least a pair of oligonucleotides, wherein at least one first oligonucleotide consists of a target-specific sequence that consists of SEQ ID NO:
- 5 joined to a 5′
promoter sequence or SEQ ID NO;
11, and wherein at least one second oligonucleotide consists of SEQ ID NO;
16 or SEQ ID NO;
38. - View Dependent Claims (18, 19, 20)
- 5 joined to a 5′
Specification