RAPID EVALUATION OF LIVE ATTENUATED AND INACTIVATED RIFT VALLEY FEVER VACCINES IN-VITRO

Document Type : Research article

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Abstract

Indirect immunofluorescence assay (IFA) was used for detection, identification and titration live attenuated RVF (7.5 log10 TCID 50 / ml) as well as inactivated alum gel RVF. Bradford's method (1976) was used to determine the amount of ug / 0.1 ml of purified reference RVF antigen (1:2000 - 1:32000) dilutions. Double sandwich ELISA was used to compare the tested inactivated RVF vaccine with a reference one has passed animal potency testing as well as to determine the amount of protein (ug / dose) of the tested inactivated RVF vaccines by using different dilutions of purified RVF antigen of known protein concentration (ug / 0.1 ml) and then a standard curve was prepared by regression analysis of the purified RVF antigen dilution of known protein concentration against its ELISA optical density (O.D.) at A450 and the protein in concentration of the reference RVF vaccine was determined by interplotting its ELISA O.D. at A405 on the RVF standard curve which reached 5 ug dose of RVF vaccine. Also, the ELISA potency ratio was determined by comparing the O.D. of tested vaccine to that of reference inactivated RVF vaccine which has passed animal potency testing and the main value of the potency ratio reached 1.00.

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