Comparison of PCR-based DNA fingerprinting techniques for the identification of species and their use for atypical isolates Free

Abstract

Four PCR-based DNA fingerprinting techniques were compared for their ability to identify at the species level a heterogeneous collection of isolates belonging to the six valid species. 16S rDNA-RFLP analysis identified all species and 16S rDNA-SSCP analysis identified almost all species. Also, isolates with unusual biochemical characteristics and/or unusual antigenic composition could be identified correctly. rRNA-intracistronic length polymorphism analysis suffered from high intraspecific variability, a limited number of fragments per profile, and small length differences between the spacers of different species. tRNA-intergenic length polymorphism analysis resulted in identification of all isolates but one, when fluorescent DNA capillary electrophoresis was used such that fragment length differences of 1 bp could be resolved. The four techniques yielded comparable results relevant to the taxonomy of . They all indicate a high degree of genetic relatedness between and , homogeneity of , distinct but clear relatedness of to the other species, a clear distinction between the two subspecies of , and a clear distinction between isolates and isolates from closely related taxa or from species which are phenotypically difficult to distinguish from . New sequence determination of the 16S rRNA gene was necessary to obtain sequences in accordance with the findings of 16S rDNA-RFLP analysis.

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1998-01-01
2024-03-29
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