Development of Enzyme Linked Immunosorbent Assay for humoral immuneresponse and infection monitoring of anthrax

Subunit vaccines or attenuated vaccines applied against Anthrax disease increase the immune systems' antibody against the PA4 which has a critical role as a toxin of Bacillus anthracis . The cornerstone of a vaccine showing its protection and efficacy in curbing the disease is the power of antibody applied against the PA4 however determination of the subjects’ exposure to the microbe or vaccine is not possible with the available Elisa kits. Material and Methods: Herein, the ELISA was developed using PA4 and LF to detect total antibody or IgG class antibody in serum. Besides, the level of anti-LF antibodies were determined as a complementary test to measure variance in antibody titers associated with vaccination or infection that leading to early detection of anthrax in livestock. Then Elisa kit was validated by checking its linearity, accuracy, precision, Quantification Limit (LOQ), Detection Limit (LOD) and specificity. The results show that we developed high-quality ELISA that can be used to test immunogenicity of vaccines and infection in mice. We tried to develop the Anti- PA4 ELISA and conduct the validation studies to evaluate the response of the antibody to the Anthrax vaccine and distinction between disease and vaccination in mice.