Development of an indirect ELISA for detection of antibody against peste des petits ruminants virus in sheep

Abstract

Background: Emergence of exotic viral disease like peste des petits ruminants(PPR) possesses a great threat towards successful goat and sheep farming in Bangladesh. Implementation of effective control measures against PPR requires a proper and rapid diagnosis of the disease. Various diagnostic techniques are available for diagnosis of PPR in goats and sheep in Bangladesh. Among these, ELISA is an important technique to diagnose the disease as well as for seromonitoring. However, the ELISA kits that are being used in the country are mostly imported. This study was undertaken to develop an in-house indirect ELISA using local PPR virus isolate as an antigen.Methods: A local isolate of PPR virus (PPRV) was used as an antigen to develop an indirect ELISA kit. Standardization of the optimum antibody, conjugate and virus antigen concentration was performed using chequer-board titration with different dilutions of the reagents. Finally, the developed indirect ELISA kit was used in a pilot study to detect anti-PPRV antibodies in sheep sera samples.Results: A polyclonal antibody based indirect ELISA was developed successfully for the detection of PPRV antibodies. The developed ELISA could detect anti-PPRV antibody in sheep sera sample during our pilot field trial.Conclusions: After robust field validation, the indirect ELISA can be used to determine the antibody titer against PPRV in the field as to monitor the seroconversion after vaccination.