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Development of probes for Brevibacterium spp.


Brevibacteria include species with both biotechnological and clinical significance. In biotechnology they contribute considerably to cheese ripening. They also have been encountered increasingly in humans as opportunistic pathogens and have been isolated from wounds, blood, an ear discharge, and from patients with otitis, osteomyelytis, peritonitis, septicemia, and groin abscess (Funke et al., 1997; Wauters et al., 2001).

The purpose of this study was to develop genus specific primers for Brevibacterium spp. in order to identify members of the genus Brevibacterium by a genotypic method.
16S sequences of all Brevibacterium type strains, additional Brevibacterium strains and 16S sequences of closely related species (negative control-strains) were aligned. Regions of similarity in Brevibacterium species were identified visually ensuring that the corresponding regions in the other species were dissimilar. These conserved regions were used to design genus-specific primers. The primers gave a fragment of approximately 236bp. when a PCR was run on Brevibacterium spp. while they gave no band with a non-Brevibacterium strain.

Brevibacteria belong to the suborder Micrococcinae. The identification of this suborder includes, among others, the determination of the type of peptidoglycan, menaquinones, and fatty acids in the cell wall. Some of these analyses are labour-intensive, which means that only few isolates can be analysed. However, accurate and reliable identification can be achieved by the 16S primers described in this study.

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