Development of an enzyme-linked immunosorbent assay using recombinant protein antigen for the diagnosis of Chikungunya virus.

We describe here the development of in-house enzyme linked immunosorbent assay (ELISA) for diagnostic Chikungunya virus (CHIKV) infection using a recombinant protein of CHIKV. Recombinant protein gene was designed based on 154 sequences and we used computational methods to predict the structure and antigenic potential.

 For the prediction of confirmation, the Monkey Recombinant Proteins gene coding for a recombinant protein CHIKV (rCHIKVp) has been synthesized and expressed in a prokaryotic system. Subsequently, the protein was purified by affinity chromatography and used as an antigen in an indirect ELISA. We present data on production optimization and preparation of the recombinant antigen ELISA to detect IgG against CHIKV in human sera.

Development of an enzyme-linked immunosorbent assay for viral infections in horses latex using recombinant E2 protein as an antigen.

Gum virus causes fever, skin eruptions, and limb edema in horses. For high-throughput and time-saving method for serodiagnosis, we explored Other Recombinant Proteins the immunogenic viral antigen sap, and establish an enzyme-linked immunosorbent assay (ELISA) using recombinant proteins. Western blot analysis using serum from infected horses show a strong reaction with the viral antigen of about 46 kDa in accordance with E1 or E2 glycoproteins.

 Recombinant E2 (RE2) proteins reacted more strongly with sera infected horses than re1 protein in both Western blotting and ELISA. In ELISA using protein Re2 (RE2-ELISA), for all horses experimentally infected with the virus latex (n = 7), the optical density (OD) exceeds the cutoff value at 14 days post-infection. BPO in five of the nine horses vaccinated also slightly exceeded the cutoff value after vaccination.

 Among the naturally infected horses (n = 28), 24 seronegative in acute sera, which turned out to be seropositive in sera recovered. During four seropositive horses in acute sera, serial dilution method endpoint with paired sera detect ≥4-fold increase in titer. In conclusion, we established RE2-ELISA can detect antibodies to the virus after infection experimental latex horse and nature; This should be useful in the diagnosis and surveillance of viral infections sap.