A novel development of Gold Nano-based indirect ELISA for sero-diagnosis of Babesiacanis.

Abstract

Innovative diagnostic techniques were used nanogold to increase the sensitivity and specificity of the original techniques of ELISA (Enzyme Linked Immunosorbent Assay); our mission production of novel kits for the diagnosis of the most common blood parasites in dogs, Babesia canis (B. canis). The protozoan antigens were formalin-killing cells. The production of antisera was performed on rats. A chemical reduction method was used to prepare gold nanoparticles. The nanoparticles synthesized were analyzed using transmission electron microscopy (TEM). The minimum antigen levels and tested sera dilutions were assessed via the checkerboard calibration; ELISA and nano-ELISA were also done. The parameters of ELISA were calculated; for instance, sensitivity and specificity, disease prevalence of antigen, positive and negative predictive values. The mean optical density (OD) of the measurements was calculated for the experimental and the control sera. The image of the synthesized gold nanoparticles (Au-NPs) by transmission electron microscopic (TEM) was rounded with a mean diameter ranging from 7.92 to 12.8 nm. The Receiver Operating Characteristic (ROC) curve for the Nano-ELISA assay is assessed against the microscopic blood examination, which serves as the gold standard. A comparison between ROC curve for the indirect ELISA assay and the gold standard of microscopic blood examination was done. In conclusion a Nanobased ELISA was the most sensitive and specific for the diagnosis of babesiosis in dogs.