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We read the recent article by Ibrahim, Gerner-Smidt, and Sjöstedt (2) with great interest. Ibrahim et al. describe a method for discrimination between closely related pathogenic bacteria based upon restriction fragment length polymorphism (RFLP) analysis of ribosomal operons. In short, they demonstrate the application of this technique to the discrimination of DNA groups 1, 2, 3, and 13 of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex, as well as to the three species of Francisella. However, the limitations of this method for taxonomic analysis must be considered. Species identification by this technique would not be possible for some bacterial genera, for instance, Haemophilus influenzae, one of the several organisms included in the study. Previously when we used the same procedure, namely, RFLP analysis with HaeIII of PCR-amplified ribosomal operons (which we termed PCR-ribotyping [3]), we observed at least 50 distinct RFLP patterns among the predominantly nonencapsulated H. influenzae in our population. While the great variation in spacer regions within the ribosomal operons makes this technique ideal for strain differentiation, the method excludes the ribosomal operon as a target for discrimination between Haemophilus species. Hence, prior knowledge of intraspecies homogeneity of the rRNA spacers would be necessary if this method were to be used for taxonomic purposes. Not only does Acinetobacter fulfil this requirement, but also all the rRNA operons in the genome appear to be identical (1). H. influenzae and a number of other organisms differ in both of these aspects (1).

journal of clinical microbiology

Amplification and restriction endonuclease digestion of a large fragment of genes coding for rRNA as a rapid method for discrimination of closely related pathogenic bacteria