Diagnosis of Clostridium Septicum Using Polymerase Chain Reaction (PCR)



Objective: Identification and confirmation of Clostridium septicum in isolated Clostridia from sheep-dung samples by PCR. Design: Laboratory study. Samples: Twenty eight Clostridia were isolated from 100 sheep-dung of Urmia. Procedure: Sheep-dung samples were collected and Clostridia isolated according to microbial tests. the DNA of isolates were extracted and used for PCR. PCR was performed by using designed primer for hemolysin gene (alpha toxin) of Cl.Septicurqx. A vaccine strain of Cl. Septicum and Cl.pefringenes types B, C and D were used as positive and negative control, respectively. Results: Six out of 28 isolates and also vaccine strain showed 270 bp band on agarose gel electrophoresis, suggesting conserved segment for hemolysin in Cl.septicutn. On the other hand, other isolates such as Clostridium fallax, Cl.perfringenes, Cl.novey B, Cl. bife rmentnas, Cl.c arnis, CI. Subterminale, Cl.rummosum, Cl. innoccum were negative. Conclusion: Since the DNA fragment of 270-bp was not amplified for Cl.perfringens, Cl.novyi, Cl.fallax' Cl. innoccum, Cl. carnis, Cl. subtenninale, Cl.bifermentance and Cl.ramusum, thts condition confirmed specificity of this primer. Hence, PCR can be useful for rapid detection or identification of Cl.septicum in clinical or environmental samples.