Jianing Zhang*, Jihong Rao*, ShuMin Ge*, #, Li-Ying Zhang**
*School of Life Science and Technology, ChangChun University of Science and Technology, ChangChun, JiLin, 130022, China - **College of Animal technology science, Jilin University,130062, China
An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) for detecting Enterovirus 71 (EV71) was developed with a monoclonal antibody (McAb)-capturing virus, while the rabbit anti-BVDV serum was used as the second antibody to identify the virus. Working conditions of AC-ELISA were optimised and its capabilities were evaluated. The results showed that the optimum working concentration of McAb was 5 μg /mL and that of the rabbit anti-EV71 serum was 10 μg /mL. The dilution of conjugate was 1:10 000, and the reaction time was one hour. Two blockings achieved the best effect. The first was 1% BSA after the 96 plates were coated with McAb, and the second was 1% gelatine after the detected antigen reacted with the coated McAb. Twenty-five EV71 samples and 24 healthy serum samples from cattle were detected in parallel by the AC-ELISA virus isolation and PCR, respectively. The accordant rate was higher. These findings suggest that the AC-ELISA has good repeatability and can be used for the rapid detection of EV71.
Antigen capture ELISA, monoclonal antibody, Enterovirus 71.
10.19193/0393-6384_2020_1_54