Isolation of Vi antigen from Salmonella Typhi Ty2 and its animal test for preparation of vaccine
Subject Areas : Molecular MicrobiologyMajid Moghbeli 1 , Ali Akbar Shaebani 2 , Elaheh AkbariFar 3
1 - Department of Microbiology, Islamic Azad university, Damghan Branch, Damghan, Iran
2 - Departmnt of Microbiology School of Public Health Semnan University, Semnan, Iran
3 - Department of Microbiology, Islamic Azad University, Qom Branch, Qom, Iran
Keywords: Vaccine, Salmonella typhi Ty2, Typhoid, Vi antigen,
Abstract :
Background and objective: Typhoid is a worldwide disease. Salmonella Typhi is the most common agent causing typhoid. It is difficult to treat typhoid due to antibiotics multiple resistance and the bacterium is menace for future epidemic. Vaccination is the most successful method against this diseases. The aim of this study is isolation of Vi antigen from Salmonella Typhi Ty2 and evaluation of it immunogenicity in animal model. Material and methods: In this research, Salmonella Typhi Ty2 was cultured in Muller Hinton Agar Medium. After harvesting the cells and to suspention them into physiologic serum, polysaccharide capsule was extracted by precipitation with cetavlone. A mount of Nucleic acid in the sample were tested with Spectrophotometere at 260 nm and amount of protein were investigated with Bradford method and SDS-PAGE and the Immunodiffusion method were used to study of isolated Vi antigen. In final, immunogenesity of Vi antigen was tested in mouse. Result: Amount of Nucleic acid and protein in the isolated sample were 0.08 µg/ml and 2.59µg respectively. In the SDS-PAGE, smear band of the Standard Vi was similar to isolated sample completely. The precipitation band in the Immunodiffusion method was confirmed to presence Vi antigen. In the study of Vi antigen on the mouse, 80 percent immunity was been created. Also injection in Rabbit showed that LPS level in the sample is not cause much fever. Conclusion: With regard to non producing the Human Vaccine of Typhoid in Iran, to be able use of this method for isolation of Vi antigen and introduce it as a vaccine candidate.
