GENERATION OF SPECIFIC PROBES FOR TARGET NUCLEOTIDE SEQUENCES

A method for generating DNA probes specific for an organism and capable of distinguishing in a non-empirical manner between species. The method comprises amplifying, using a pair of oligonucleotide primers, a variable region of the genome of a number of phylogenetically related organisms to, or of a...

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Bibliographische Detailangaben
Hauptverfasser: RICHARD POWELL, THOMAS GERARD BARRY, BERNARD FRANCIS XAVIER GANNON
Format: Patent
Sprache:eng
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Beschreibung
Zusammenfassung:A method for generating DNA probes specific for an organism and capable of distinguishing in a non-empirical manner between species. The method comprises amplifying, using a pair of oligonucleotide primers, a variable region of the genome of a number of phylogenetically related organisms to, or of a number of organisms suspected of being present in a given sample containing, said organism to be identified and having said variable region in its genome, at least one of said primers corresponding to a DNA sequence known or suspected of being conserved in said organisms, determining the sequence of the amplified region, selecting said sequence or a portion thereof to generate said probe specific for said organism to be identified by comparison with said other amplified regions and defining the hybridization conditions required to obtain a specific signal based on the precise nucleotide sequence of the selected probe. Specific probes are disclosed for a variety of species including Aeromonas hydrophila, Aeromonas salmonicida, Clostridium difficile, Mycobacterium bovis, Mycobacterium tuberculosis and Salmonella typhimurium.