Institut Pasteur

Stephen Gordon

Stewart T Cole

Roland Buchrieser-Brosch

Alain Billault

Roland Brosch

Thierry Garnier

Karin Eiglmeier

Glyn Hewinson

A method for detectinginvolves contacting a biological sample with the product of expression of at least one ORF contained in a polynucleotide present inand absent inBCG; and detecting whether an immunological complex is formed between the product thereof and antibodies contained in the biological sample, wherein the presence of a complex indicates an infection with

Method for isolating a polynucleotide of interest from the genome of a mycobacterium using a BAC-based DNA library: application to the detection of mycobacteria

The present invention is the identification of a nucleotide sequence which make it possible in particular to distinguish an infection resulting from the vast majority ofstrains from an infection resulting fromBCG. The subject of the present invention is also a method for detecting the sequences in question by the products of expression of these sequences and the kits for carrying out these methods. Finally, the subject of the present invention is novel vaccines.

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The present invention is directed to a method for isolating a polynucleotide of interest that is present or is expressed in a genome of a first mycobacterium strain and that is absent or altered in a genome of a second mycobacterium strain which is different from the first mycobacterium strain using a bacterial artificial chromosome (BAC) vector. The invention further relates to a polynucleotide isolated by this method and recombinant BAC vector used in this method. In addition the present invention comprises method and kit for detecting the presence of a mycobacteria in a biological sample.

A method for isolating a polynucleotide of interest that is present in the genome of a first mycobacterium strain and/or is expressed by the first mycobacterium strain, where the polynucleotide of interest is also absent or altered in the genome of a second mycobacterium strain and/or is not expressed in the second mycobacterium. The method includes (a) contacting the genomic DNA of the first mycobacterium strain under hybridizing conditions with the DNA of a least one clone that belongs to a bacterial artificial chromosome (BAC) genomic DNA library of the second mycobacterium strain, and (b) isolating the polynucleotide of interest that does not form a hybrid with the DNA of the second mycobacterium strain. This invention further pertains to astrain H37Rv genomic DNA library, as well as aBCG strain Pasteur genomic DNA library, and the recombinant BAC vectors that belong to those genomic DNA libraries. This invention also relates to mycobacterial nucleic acids, and methods and kits for using these nucleic acids to detect mycobacteria in a biological sample.

Method for isolating a polynucleotide of interest from the genome of a mycobacterium using a BAC-based DNA library, application to the detection of mycobacteria

A method for isolating a polynucleotide of interest that is present in the genome of a first mycobacterium strain and/or is expressed by the first mycobacterium strain, where the polynucleotide of interest is also absent or altered in the genome of a second mycobacterium strain and/or is not expressed in the second mycobacterium. The method includes (a) contacting the genomic DNA of the first mycobacterium strain under hybridizing conditions with the DNA of a least one clone that belongs to a bacterial artificial chromosome (BAC) genomic DNA library of the second mycobacterium strain, and (b) isolating the polynucleotide of interest that does not form a hybrid with the DNA of the second mycobacterium strain. This invention further pertains to a strain H37Rv genomic DNA library, as well as a BCG strain Pasteur genomic DNA library, and the recombinant BAC vectors that belong to those genomic DNA libraries. This invention also relates to mycobacterial nucleic acids, and methods and kits for using these nucleic acids to detect mycobacteria in a biological sample.

Method for isolating a polynucleotide of interest from the genome of a mycobacterium using a bac-based DNA library: application to the detection of mycobacteria

The present invention relates to a method for isolating a polynucleotide of interest that is present in the genome of a first mycobacterium strain and/or is expressed by the first mycobacterium strain, where the polynucleotide of interest is also absent or altered in the genome of a second mycobacterium strain and/or is not expressed in the second mycobacterium. The method comprises: (a) contacting the genomic DNA of the first mycobacterium strain under hybridizing conditions with the DNA of a least one clone that belongs to a bacterial artificial chromosome (BAC) genomic DNA library of the second mycobacterium strain, and (b) isolating the polynucleotide of interest that does not form a hybrid with the DNA of the second mycobacterium strain. This invention further pertains to a,strain H37Rv genomic DNA library, as well as a,BCG strain Pasteur genomic DNA library, and the recombinant BAC vectors that belong to those genomic DNA libraries. This invention also relates to a method, as well as a kit, for detecting a nucleic acid of a mycobacterium in a biological sample.

Method for isolating a polynucleotide of interest from the genome of a mycobacterium using a BAC-based DNA library application to the detection of mycobacteria

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Paris, France

Stephen Gordon