Rapid detection and identification of Clostridium chauvoei by PCR based onflagellin gene sequence

We developed a one-step polymerase chain reaction (PCR) system that specifi cally detects Clostridium chauvoei. Oligonucleotide primers were designed t o amplify a 516-bp fragment of the structural flagellin gene. The specifici ty of the PCR was investigated by analyzing 59 strains of clostridia, and s even strain of other genera. A 516-bp fragment could be amplified from all the C. chauvoei strains tested, and no amplification was observed by using DNAs from the other strains tested, including Clostridium septicum. Similar ly, this PCR-based method specifically detected C. chauvoei DNA sequences i n samples of muscle and exudate of obtained from mice within 12 h of inocul ation. In tests using samples of muscle or liver, the limit of detection wa s about 200 organisms per reaction. These results suggest that the one-step PCR system may be useful for direct detection and identification of C. cha uvoei in clinical specimens. (C) 2001 Elsevier Science B.V. All rights rese rved.