Developing ELISA test formula to detect JEV infection in amplifying host (avian and pig)

 Vector-borne and zoonotic diseases occupy the top position as infectious diseases that have emerged in humans in the past few decades. One of the vector-mediated zoonotic diseases is Japanese encephalitis (JE). JE disease is caused by JE virus

The JE virus occurs naturally in birds from the Ardeidae family and the virus is transmitted from birds to birds through mosquito intermediaries. JE virus can be transferred to pigs which are amplifiers, the virus is spread to pigs and then transmitted to other animals or humans. In humans, this virus causes serious acute encephalitis, and humans are the dead-end hosts of the JE virus. Because the incidence of JE in humans is very closely related to the presence of viruses in animals. Surveillance of JE in animals needs to be done regularly to anticipate the explosion of the JE case in humans. The obstacle faced in carrying out this antibody serosurvey is the high cost of commercial ELISA kits for animals. Even ELISA kits for chickens and ducks are no longer produced. 

We have successfully formulated the Indirect ELISA test to detect antibodies against JE using synthetic JE antigens. The JE antigen is coated overnight on the ELISA microplate, then it is blocked with Skim milk, the serum to be tested is added to the microplate, following with secondary antibodies labeled HRP, the antigen and antibody complex are tracked using a TMB substrate. With this formula, antibodies can be tested for JE on swine, chicken, and duck without being dependent on an expensive  commercial ELISA Kit.