Conventional and molecular identification of vaccine strain of Clostridium septicum

Introduction and Objectives: Genus Clostridium are ubiquitous which are cause disease in human and animal. Disease caused by Clostridium species have been existed from years ago in Iran widespread. For prevention require effective vaccines. So, toxigenic strains are needed. The purpose of this study is characterization of vaccine strain of using microbiological and biochemical properties, molecular approach and toxicity titration. Materials and Methods: Vaccine strain (CN913) was cultured anaerobically in liver broth for 24 h. After growth, this strain was characterized using microbiological (including Gram staining, culture on Blood agar and LB agar and motility) and biochemical tests (including Fermentation of sugars, lecithinase, gelatinase, Indol, and catalase). Then genomic DNA was extracted using phenol and chloroform extraction method. The target gene was amplified through PCR by specific primer which was taken from gene bank and designed using oligo software. Finally, Toxicity titration was performed using injection intravenously into white mice per dilution. The inoculated mice were observed and titer was calculated. Result: C. septicum colonies were smooth and slightly translucent colonies surrounded by beta haemolysis. In Gram staining was observed gram positive straight or curved rod shape. The other microbiological and biochemical properties of vaccine strain are also found similar to those of C. Septicum. On the other hand, Vaccine strain was confirmed using PCR approach. The titer of lethal toxin produced was 1:50 per ml. Conclusion: The result showed that vaccine strain is confirmed with reliable references and could be used as vaccine strains.

کلیدواژه ها:

Clostridium septicum ، PCR ، Diagnosis ، toxin-alpha