Polymerase chain reaction primers and probes for Mycobacterium tuberculosis
AssigneeRutgers, The State University of New Jersey
InventorsDavid Alland, Soumitesh Chakravorty
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Abstract
The present invention relates to novel primers and sloppy molecular beacon and molecular beacon probes for amplifying segments from different genes in Mycobacterium tuberculosis for identifying the presence of M.tb DNA and/or resistance to anti-tuberculosis drugs.
Claims8
An oligonucleotide set for amplifying a portion of a Mycobacterium tuberculosis katG gene, comprising,
- a pair of forward and reverse primers specific for said portion, wherein the forward primer comprises the sequence of SEQ ID NO:
- 7, 8, or 9 and the reverse primer comprises the sequence of SEQ ID NO:
- 10 or 11 and
- an isolated nucleic acid probe consisting of the sequence of SEQ ID NO:
- 56, wherein the nucleic acid probe is labeled with a fluorophore and a quencher at its two ends respectively.
An isolated nucleic acid probe consisting of the sequence of SEQ ID NO:
- 56 wherein the nucleic acid probe is labeled with a fluorophore and a quencher at its two ends respectively.
A method for detecting presence of Mycobacterium tuberculosis in a test sample, comprising contacting the test sample with a pair of forward and reverse primers specific for a portion of a katG gene, wherein the forward primer comprising the sequence of SEQ ID No:
- 7, 8, or 9 and the reverse primer comprises the sequence of SEQ ID No:
- 10 or 11, under conditions conductive to amplifying reaction to yield a first amplicon, hybridizing the first amplicon with a probe consisting of the sequence of SEQ ID No:
- 56, and detecting Mycobacterium tuberculosis based upon the hybridization.
A method for detecting presence of Mycobacterium tuberculosis in a test sample, comprising contacting the test sample with a pair of forward and reverse primers specific for a portion of a katG gene, wherein the forward primer comprising the sequence of SEQ ID No:
- 7, 8, or 9 and the reverse primer comprises the sequence of SEQ ID No:
- 10 or 11, under conditions conductive to amplifying reaction to yield a first amplicon, hybridizing the first amplicon with a molecular beacon probe consisting of the sequence of SEQ ID No:
- 56 and labels of a fluorophore and a quencher at the two ends of the sequence, and detecting Mycobacterium tuberculosis based upon the hybridization.
Dependencies are inferred from the claim text.