purification and preparation of specific antibody against 146 S antigen of FMD virus current serotypes in Iran.
2010
Zibaei, Saeid | Mirjalili | Fathiyeh najafi | Hemati | Khorasani
Foot and mouth disease (FMD) is the most contagious disease of mammals and has a great potential for causing severe economic loss in susceptible cloven-hoofed animals. FMDV is a member of the genus aphthovirus in the family Picornaviridae. There are seven serotypes of FMD virus, namely, O, A, C, SAT 1, SAT 2, SAT 3 and Asia 1. Infection with one serotype does not confer immunity against another. There are three serotypes of FMD virus in Iran (A, O, Asia1) and produced vaccine against the serotypes. Because of speed of contagion disease, detection rate of FMDV is very important. In direct sandwich ELISA for FMD antigen detection and typing is applied as a routine in most FMD diagnostic laboratories. At present, in direct sandwich ELISA kit not produced in Iran. Purification 146 S antigen of FMD virus and produced specific antibody against the antigen in guinea pigs and rabbits are the first step of produced of the ELISA kit. In these study 146s particles of FMDV serotypes A,O and Asia1 purified by using of sucrose gradient ( 20%-50% ) .Dot blot , in direct ELISA and SDS page tests are used for approved of purification. The specific antigens were produced based on protocol in guinea pigs and rabbits by inoculation 25 g of 146s particles of FMDV with complete and subsequently incomplete Freund s adjuvant .Rapid agglutination, dot blot and in direct ELISA tests were used for approved of produced specific antigens. For purification of specific antibodies against 146s particles used ion Exchange chromatography after precipitated with 40% ammonium sulfate. Dot blot , in direct ELISA and SDS page tests are used for approved of purified antibodies for serotypes A,O and Asia1 and indirect sandwich ELISA and western blot were used for serotype O. According to the results purification of 146s particles of FMDV serotypes A, O and Asia1and production of specific antibody against those were approved.
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