Submitted to: Journal of Clinical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/8/2002
Publication Date: 10/1/2002
Citation: Boonchit, S., Xuan, X., Yokoyama, N., Goff, W.L., Wagner, G.G., Igarashi, I. 2002. Evaluation of an enzyme-linked immunosorbent assay with recombinant rhoptry-associated protein-1 antigen against babesia bovis for the detection of specific antibodies in cattle. Journal of Clinical Microbiology. 40:3771-3775. Interpretive Summary: Babesia bovis is a tick-transmitted microorganism, causing disease in cattle throughout much of the tropical and subtropical world. Other related species of Babesia also cause disease, but each in a slightly different way. In addition, more than one species occurs many areas. Thus, assays for differentiating between different babesial species are very important. However, these specific assays are not available in formats that allow for easy and quick diagnosis. In this study, we detail the use of a recently characterized protein associated with Babesia bovis to detect antibody present in infected cattle. The assay was able to discriminate between Babesia bovis infected cattle and cattle infected with other pathogenic babesial species.
Technical Abstract: The gene encoding Babesia bovis rhoptry-associated protein-1 (RAP-1) was used to develop an enzyme-linked immunosorbent assay (ELISA) to measure specific antibodies against B. bovis. The B. bovis RAP-1 gene was subcloned into a baculovirus transfer vector and the RAP-1 protein was expressed in insect cells infected with a recombinant baculovirus. The recombinant B. bovis RAP-1 of 65 kDa was detected with anti-RAP-1 mouse serum by Western blotting, and this recombinant RAP-1 was used as an antigen in the ELISA. The ELISA was able to differentiate between B. bovis-infected sera and B. bigemina infected sera or noninfected normal bovine sera. The results demonstrate that the recombinant RAP-1 expressed in insect cells might be a useful antigen for the detection of antibodies to B. bovis.