Compare of two methods of direct PCR and PCR with DNA extraction in Clostridium Perfringens typing

    Polymerase chain reaction (PCR) is a suitable method for detecting pathogenic agents and first step in this process is DNA extraction. DNA extraction enumerate as a long stage in molecular genetic researches that is longer than total PCR process in some cases. The aim of this research is use of direct PCR (without DNA extraction) for different typing of Clostridium perfringens. In this study direct PCR and also PCR by extracted DNA was implemented on the A, B, C and D types of Clostridium perfringens. For this purpose four pairs of specific primers that were complementary with parts of the genes encoding α, β, ε and ι toxins of this bacterium were used. In both direct PCR and PCR by extracted DNA, 196, 324 and 655 bp fragments were amplified that are dependent to α, β and ε toxins respectively. Upshot different strains of this bacterium were identified based on amplified fragment length in PCR. The result showed that success of PCR is simple from colony directly and quality of created bands is the same in both methods. Therefore, use of this method is recommended because the direct PCR reduce time, steps and cost of PCR and provide possibility of testing many samples in shorter times provides.