Evaluation of recombinant Babesia gibsoni thrombospondin-related adhesive protein (BgTRAP) for the sero-diagnosis of canine babesiosis

Exp Parasitol. 2023 Nov:254:108621. doi: 10.1016/j.exppara.2023.108621. Epub 2023 Sep 16.

Abstract

Canine babesiosis, caused by Babesia gibsoni is one of the most significant tick-borne illnesses across the world. Light microscopy as well as polymerase chain reaction may fail in the diagnosis of disease when the level of parasitaemia is very low during subclinical and chronic cases. The serological techniques using a recombinant protein will be useful for the accurate and sensitive surveillance of the disease, especially in chronic cases. The present study describes the evaluation of recombinant N-terminal B. gibsoni Thrombospondin-related adhesive protein (BgTRAP) based indirect ELISA for the sero-diagnosis of B. gibsoni infection in dogs. A partial N-terminal BgTRAP gene (870 bp) of B. gibsoni, was expressed in Escherichia coli using a pET32a (+) vector. The recombinant BgTRAP based indirect ELISA was compared with the PCR targeting the same gene. A sensitivity and a specificity of 84% and 73.33% were observed in the indirect ELISA. The accuracy, positive predictive value and negative predictive value were 78.18%, 72.30%, 84.60% respectively. The rBgTRAP antigen did not show any cross-reactivity with sera from dogs infected with common helminth parasites viz. Ancylostoma caninum, Dirofilaria immitis, D. repens, Spirometra spp., Toxocara canis and haemoparasites like Trypanosoma evansi, Babesia vogeli, Hepatozoon canis and Ehrlichia canis.

Keywords: B. gibsoni; BgTRAP; Dogs; Indirect-ELISA; PCR; Prokaryotic expression.