Conventional and molecular characterization of Clostridium perfringens type B vaccine strain

Introduction and Objectives: Clostridium perfringens is a spore-forming gram-positive bacterium which is found in the intestines of humans and animals. The best way to control these diseases is through vaccination. The purpose of this study is characterization of vaccine strain, using microbiological and biochemical properties, molecular approach and toxicity titration. Materials and Methods: Vaccine strain (CN228) was cultured anaerobically in liver broth for 24 h. Afterwards, characterization of strain was done 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 retrieved from gene bank and designed using OLIGO software. Finally, MLD of toxin was performed using intravenous injection into white mice tail vein. The challenged mice were observed daily over 3 days to determine toxin activity. Result: C. perfringens colonies are smooth, round, and grayish which are surrounded by a double zone of hemolysis. 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. perfringens type B. On the other hand, Vaccine strain was confirmed using PCR analysis. Conclusion: The result showed that vaccine strain is confirmed with reliable references and could be used as vaccine strain.

کلیدواژه ها:

Clostridium perfringens type B ، PCR ، Diagnosis ، toxin