Application of molecular methods for routine identification of acetic acid bacteria.

Recently many new species of Acetic acid Bacteria have been described. The description and identification as new species was based on molecular techniques (sequencing of the 16S rRNA gene, DNA base ratio (% GC) determinations and DNA-DNA hybridisation) and phenotypic characterization. In the present paper, we propose a fast and reliable method for the identification most of the species currently described based on the RFLP-PCR of the 16S rRNA. According to the proposed protocol, 1 species can be identified with the use of a single enzyme, 13 with a combination of 2 enzymes, 2 species with a combination of 3 enzymes, 2 with a combination of 4 enzymes. To differentiate 5 more species RFLP-PCR of the ITS was also needed, after using 3 enzymes. Finally, a pair of species (Acetobacter pasteurianus and Acetobacter pomorum) could not be distinguished with the proposed method. However, doubts can be raised about their differentiation as separate species. Keeping these limitations in mind, the method is fast and reliable, allowing the processing of large number of samples in relatively short periods of time (less than 24 h after the isolation).