Genomic analysis method for the detection and identification of mycobacteria
Inventor: Stewart COLE, Roland BUCHRIESER-BROSCH et al.
Description of invention:
Over 1.7 billion people worldwide--a third of the world’s population--harbor dormant tuberculosis infections. According to the World Health Organization (WHO), there will be more than 8 million new cases of active tuberculosis diagnosed this year, and over 2.9 million tuberculosis-related deaths. WHO estimates that nearly 1 billion tuberculosis tests will be performed this year and that testing will increase l0-l4% annually.Current diagnostic tools for tuberculosis are at times inadequate and time consuming, some requiring weeks before a diagnosis is obtained.
Researchers at the Institut Pasteur have designed a convenient method for detection and identification of type-specific genomic variation between distinct mycobacterium strains. The method is based on a defined bacterial artificial chromosome (BAC) library including the whole genome of a reference strain, which is hybridized and compared with DNA extracted from biological samples. The detection method focuses on a strain-specific polynucleotide used for distinction between the mycobacteria strains for diagnostic purposes. The invention further relates to purified polynucleotides isolated by this method and recombinant BAC vectors used in this method.
The recombinant BAC vectors can be used to detect mycobacterial nucleic acids (genomic DNA, cDNA or mRNA) in biological samples. The polynucleotides identified are useful as probes or primers for detecting a given mycobacterium of interest. The BAC cloning system is advantageous over mycobacterial DNA libraries constructed in cosmid clones because the F-plasmid is present in only one or a maximum of two copies per cell, reducing the potential for recombination between DNA fragments, and more importantly, avoiding the lethal overexpression of cloned bacterial genes. This invention represents an important tool in the global effort to combat tuberculosis.
Recent pertinent publications:
* Brosch R et al. Infect Immun 1998 May;66(5):2221-9
* S. V. Gordon […], S.T. Cole, R. Brosch et al. Nature, Vol 393, 11 june 1998
* S.T. Cole, Molecular Microbiology, 1999, 32(3), 643-655
Various patents pending
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