Rapid identification and differentiation of pathogenic clostridia in gas gangrene by polymerase chain reaction based on the 16S-23S rDNA spacer region

In cattle, sheep, and other ruminants, clostridial myonecrosis (gas gangren e) is mostly caused by Clostridium chauvoei, C septicum, C novyi and C sord ellii. A polymerase chain reaction (PCR) system using common primers design ed from multiple alignment of the 16S rRNA and 23S rRNA genes of Clostridiu m species was developed to identify pathogenic clostridia. The PCR was perf ormed with total DNA from 26 strains which included seven different Clostri dia species. These bacteria were differentiated at species level by the dif ferent pen product patterns. To characterise the 16S-23S rDNA spacer region s of these clostridia further, most PCR products of these bacteria were seq uenced. The smallest PCR products of each bacterium represented the fundame ntal 16S-23S rDNA spacer region; larger PCR products of each bacterium were caused by insertion sequences, i.e. tRNA gene sequences. The authors' obse rvations indicate that the PCR patterns of the 16S-23S rDNA spacer regions have the potential to be used as an identification marker of pathogenic clo stridia in gas gangrene. (C) 2000 Harcourt Publishers Ltd.