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.
Claims11
An isolated nucleic acid probe for detecting drug-resistant M.tb comprising a probe sequence that is at least 85% identical to SEQ ID No:
- 56 in its entirety, wherein the probe is 38 to 50 nucleotides in length, and wherein the probe is labeled with a fluorophore and a quencher at its two ends respectively.
An oligonucleotide set for amplifying a portion of an M. tuberculosis katG gene, comprising:
- i. a first pair of forward and reverse primers specific for a first portion of the katG gene, wherein each primer has a sequence that is at least 85% identical to an oligonucleotide sequence selected from SEQ ID Nos:
- 7-11;
- and
- ii. an isolated nucleic acid probe comprising a probe sequence that is at least 85% identical to SEQ ID No:
- 56 in its entirety, wherein the probe is 38 to 50 nucleotides in length, and wherein the probe is labeled with a fluorophore and a quencher at its two ends respectively.
A method for detecting presence of M. tuberculosis in a test sample, comprising
- contacting the test sample with a first primer pair under conditions conducive to an amplifying reaction to yield a first amplicon, and
- detecting the presence of said amplicon thereby detecting presence of Mycobacterium tuberculosis in said test sample,
- wherein the first primer pair is specific for a portion of a M. tuberculosis katG gene, and
- wherein each primer of the first primer pair has a sequence that is at least 85% identical to an oligonucleotide sequence selected from the group consisting of SEQ ID Nos:
- 7-11.
A method for detecting presence of M. tuberculosis in a test sample, comprising
- contacting the test sample with a first molecular beacon probe under conditions conducive to a hybridization reaction to yield a probe-target hybrid, and
- detecting the presence of said probe-target hybrid thereby detecting presence of Mycobacterium tuberculosis in said test sample,
- wherein the first molecular beacon probe comprises a sequence that is at least 85% identical SEQ ID No:
- 56.
Dependencies are inferred from the claim text.