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GENERATION OF SPECIFIC DNA PROBES FOR GENERA AND SPECIES DIFFERENTIATION

机译:特定物种和物种分化的特定DNA探针的产生

摘要

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.
机译:一种产生对生物体特异性的DNA探针的方法,该方法能够以非经验的方式在物种之间进行区分。所述方法包括使用一对寡核苷酸引物,扩增与给定样品中存在的多种系统发育相关生物的基因组可变区,或疑似存在于给定样品中的多种生物的基因组可变区,所述生物要鉴定并具有在其基因组中的所述可变区,至少一个所述引物对应于已知或怀疑在所述生物中保守的DNA序列,确定扩增区的序列,选择所述序列或其一部分以产生对所述特异性的探针通过与所述其他扩增区域进行比较并根据所选探针的精确核苷酸序列确定获得特定信号所需的杂交条件来鉴定该生物。公开了针对多种物种的特异性探针,包括嗜水气单胞菌,鲑鱼气单胞菌,艰难梭菌,牛分枝杆菌,结核分枝杆菌和鼠伤寒沙门氏菌。

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