A 16S rDNA-based PCR method for rapid and specific detection of Clostridium perfringens in food

Abstract

A 16S rDNA-based polymerase chain reaction (PCR) method was developed for the rapid and specific detection of Clostridium perfringens in food. The PCR primers were designed by a GenBank computer search and they are complementary only with the 16S rRNA gene of C. perfringens by sequence alignment. The PCR product is a 279 BP DNA fragment. All C. perfringens strains tested were positive in the PCR assay and all other species tested were negative, including 11 other species of Clostridium and 38 species of other common bacteria. As few as two cells of C. perfringens in pure culture were detectable. High numbers of other bacterial species did not interfere with the detection of C. perfringens. The PCR amplification required only 30 min to complete. The method can be used for detection of C.perfringens in contaminated food. Samples from 100 g of chicken drumsticks which were inoculated with 20, 200 or 2000 cells of C.perfringens were subjected to the PCR assay and all produced positive results.