Rapid detection of antibiotic resistance in mycobacterium tuberculosis
AssigneeInstitut Pasteur
InventorsBeate Heym, Stewart Cole, Douglas Young, Ying Zhang, Nadine Honore, Amalio Telenti, Thomas Bodmer
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Abstract
A process for the detection of resistance to an antibiotic in a mycobacterium comprises detecting a mutation in a gene selected from the group consisting of the katG gene or fragment thereof, the rpoB gene or fragment thereof, and the rpsI gene or fragment thereof. The process is useful for detecting in vitro the presence of nucleic acids of a Mycobacterium tuberculosis resistant to isoniazid.
Claims21
A process for the detection of a resistance to an antibiotic in a mycobacterium which comprises detecting a mutation in a gene selected from the group consisting of the katG (SEQ ID NO:45) gene or fragment thereof, the rpoB (SEQ ID NO:
- 59) gene or fragment thereof and the rpsL (SEQ ID NO:63) gene or fragment thereof.
A kit for the detection of Mycobacterium tuberculosis resistant to isoniazid, wherein the kit comprises:
- (A) a container means containing a probe labelled by a label selected from the group consisting of radioactive, enzymatic, fluorescent, and luminescent labels, comprising a nucleic acid sequence, which is a 2.5 kb EcoRV-KpnI fragment of plasmid pYZ56 or part thereof, wherein said fragment contains a BamHI cleavage site and to provide for the selectivity of the in vitro detection of a Mycobacterium tuberculosis resistant to isoniazid;
- and
- (B) a container means containing a control preparation of nucleic acid.
A nucleic acid probe for detecting Mycobacterium tuberculosis resistant to isoniazid, wherein said probe consists of a 2.5 kb EcoRV-KpnI fragment of plasmid pYZ56, wherein said fragment contains a BamHI cleavage site, or of a part of said fragment nonetheless sufficiently long to provide for the selectivity of the in vitro detection of a Mycobacterium tuberculosis resistant to isoniazid.
A kit for the in vitro of the resistance of a bacteria of a mycobacterium genus of isoniazid or its analogoues comprising:
- primers for the amplification of the DNA of the katG gene (SEQ ID NO:45) or a fragment thereof;
- reagents for amplifying said DNA;
- at least one detectably labeled probe capable of hybridizing with the amplified nucleotide sequence to be detected;
- and
- a control DNA of katG gene (SEQ ID NO:45) of a strain of said bacteria sensitive to isoniazid or a fragment thereof.
A kit for the in vitro diagnostic of the resistance of a bacteria of a mycobacterium genus to rifampicin or its analogues, characterized in that it comprises:
- primers for the amplification of the DNA of the rpoB gene (SEQ ID NO:59), the .beta.-sub-unit of the RNA polymerase of said mycobacteria, or a fragment thereof;
- reagents for amplifying said DNA;
- at least one detectably labeled probe capable of hybridizing with the amplified nucleotide sequence to be detected;
- and
- a control DNA of rpoB gene (SEQ ID NO:45) coding for the .beta.-sub-unit of the RNA polymerase of a strain of said bacteria sensitive to rifampicin, or a fragment thereof.
A kit for the in vitro diagnostic of the resistance of a bacteria of a mycobacterium genus to streptomycin or its analogues, characterized in that it comprises:
- primers for the amplification of the DNA of the rpsL gene (SEQ ID NO:63) coding for the S 12 protein of the small ribosome sub-unit or a fragment thereof;
- reagents for amplifying said DNA;
- at least one detectably labeled probe capable of hybridizing with the amplified nucleotide sequence to be detected;
- and
- a control preparation of a DNA of rpsL gene (SEQ ID NO:65) coding for the S12 protein of the small sub-unit of the ribosome (SEQ ID NO:66) of the M. tuberculosis strain sensitive to streptomycin, or a fragment thereof.
Dependencies are inferred from the claim text.