Detection of rpoB sequences of Mycobacterium tuberculosis
AssigneeGen-Probe Incorporated
InventorsMarkus T. Jucker, Steven T. Brentano, Francisco D. Delgado, Philippe Cleuziat
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Abstract
A method of detecting rpoB sequences of Mycobacterium tuberculosis present in a biological sample that includes steps of amplifying the M. tuberculosis rpoB sequence in vitro in a nucleic acid amplification mixture that includes specific disclosed primer sequences, and detecting the amplified sequences using multiple probes that provide sequence information by their specific hybridization to portions of the amplified nucleic acid. Compositions for amplifying and detecting in vitro the rpoB sequences of M. tuberculosis in a sample are disclosed.
Claims21
A method of detecting rpoB sequences of Mycobacterium tuberculosis present in a biological sample, comprising the steps of:
- providing a biological sample containing nucleic acid from M. tuberculosis comprising a rpoB sequence;
- amplifying the rpoB sequence in an in vitro nucleic acid amplification reaction mixture comprising at least one polymerase activity, and primers consisting of SEQ ID NO:2 with SEQ ID NO:
- 3,
- to produce amplified nucleic acid containing a rpoB sequence;
- optionally fragmenting the amplified nucleic acid;
- hybridizing the amplified nucleic acid to at least one detection probe consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4 that hybridizes specifically to M. tuberculosis sequences;
- and
- detecting the amplified nucleic acid hybridized to at least one detection probe by detecting a label associated with the amplified nucleic acid.
A composition for detecting a rpoB sequence of M. tuberculosis, comprising oligonucleotides consisting of SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:
- 4.
A composition for detecting a rpoB sequence of M. tuberculosis, comprising oligonucleotides consisting of SEQ ID NO:2, SEQ ID NO:3, and at least one detection probe selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4.
A kit comprising at least three oligonucleotides, wherein two oligonucleotides consist of SEQ ID NO:2 and SEQ ID NO:3, and at least one other oligonucleotide is selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:4.
Dependencies are inferred from the claim text.