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.
Claims28
An oligonucleotide set for amplifying a portion of a M. tuberculosis region selected from the group consisting of rpoB gene, gyrA gene, gyrB gene, inhA promoter, rrs gene, eis promoter, embB gene, katG gene, dosR gene, IS6110 gene, IS1081 gene, comprising, a pair of forward and reverse primers specific for said portion, wherein each primer has a sequence that is substantially identical to an oligonucleotide sequence selected from those listed in Tables 1A and 1B.
An isolated nucleic acid comprising a sequence that is substantially identical to one selected from those listed in Table 2.
A method for detecting drug resistance in M. tuberculosis, comprising
- amplifying a first nucleic acid target sequence with a first primer pair to generate a first amplicon, the first primer pair being specific for a portion of a region selected from the group consisting of rpoB gene, gyrA gene, gyrB gene, inhA promoter, rrs gene, eis promoter, embB gene, and katG gene and each primer having a sequence that is substantially identical to an oligonucleotide sequence selected from those listed in Tables 1A and 1B, and
- detecting a mutation in the first amplicon,
- wherein the presence of the mutation is indicative of the drug resistance.
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 an oligonucleotide set for amplifying a portion of a M. tuberculosis region selected from the group consisting of gyrB gene, inhA promoter, eis promoter, embB gene, katG gene, dosR gene, IS6110 gene, IS1081 gene and
- wherein each primer of the first primer pair has a sequence that is substantially identical to an oligonucleotide sequence selected from those listed in Table 1B.
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 substantially identical to one selected from those listed in Table 2.
Dependencies are inferred from the claim text.