To investigate whether 16S-23S rDNA (rDNA) spacer region length polymorphisms are suitable for the identification of , the 16S-23S rDNA intergenic spacer region lengths of 221 strains belonging to 31 species were studied by using a PCR-based method. Each species presented a specific 16S-23S pattern made of 1–8 fragments ranging from 104 to 771 bp, with the exception of the species and , which presented larger or smaller fragments. Very few species showed more than one pattern, subsp. and being the most heterogeneous species (five different patterns for eight strains). Five clinical strains that could not be identified at the species level by phenotypical tests were finally identified using this method. Discrimination between some species that showed close patterns ( equorum, ) was further achieved after digestion of the PCR products. Although it does not allow discrimination of subspecies, the use of 16S-23S spacer region length data determined by PCR-mediated amplification is suitable for the identification of the 31 tested in this study. The method is rapid, easy and may be a useful tool for the identification of species in the clinical microbiology laboratory.


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