Establishment Of 3ABC-ELISA For Foot-and-Mouth Disease Virus

Foot-and-mouth disease (FMD) is a highly contagious disease of animals which caused by Foot-and-mouth disease virus (FMDV). For the control of the disease, a clear technical system has been established: in developed countries, slaughtering of suspected and diseased animal was often adopted, which can rapidly break down the disease and be recovered to disease free status. Nevertheless, in developing countries,systematic control measure based on vaccination is usually used. Considerable effort and attention is now being directed toward the development of new methods and techniques for detection of non-structural protein(NSP)antibody which have been accepted as a reliable methods for differentiation of FMDV infected animals from vaccinated animals.Non-structure protein gene 3ABC,3AB and 3B of FMDV was expressed by baculovirus expression system. The recombinant proteins have apparent molecular weights of about 37 Ku and 26 Ku, respectively, as indicated by SDS-PAGE analysis, and showed specific immunoreactivity in Western blot. An indirect ELISA (3ABC-ELISA) was established using the purification protein 3ABC as coating antigens. The optimal coating amount was determined to be 2.5μg/Ml, the confining solution was optimalized at skimmed milk and the incubation time was 1h, the optimal incubation time of sera was 1h, the incubation time of substrate was 15min. The sensitivity show that the dilution at which there was detected of 1:1280. No cross-reaction occurred when Infectious Bovine Rhinotracheitis(IBR),Akabane Diseasa(AKD),Bovine Ephemeral Fever(BEF),Contagious Bovine Pleuropneumonia(CBPP)were tested. A total 44 sera from cattle were detected in parallel by 3ABC-ELISA and NSP-ELISA test kit of Lan Zhou Veterinary Research Institute for FMDV and test kit of YuanHeng Biotechnology, the conformance was 86.4% and 77.3%。...