Characterization of the Babesia gibsoni 12-kDa protein as a potential antigen for the serodiagnosis |
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Authors: | Youn-Kyoung Goo Honglin Jia G Oluga Aboge M Alaa Terkawi Eung-goo Lee Junya Yamagishi Yoshifumi Nishikawa Hyung-Kwan Jang Fujiko Sunaga Kazuhiko Namikawa Kozo Fujisaki Xuenan Xuan |
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Institution: | 1. Faculty of Agricultural Sciences, University of Camagüey, Ignacio Agramonte Loynáz. Cuba. Carretera de Circunvalación Km 5 1/2, Camagüey 74 650, Cuba;2. Department of Veterinary Medicine, Federal University of Parana, Rua dos Funcionários, 1540 - Laboratório de Doenças Parasitárias, HV Juvevê, 80035-050 Curitiba, PR, Brasil;3. Department of Veterinary Science, University of Kentucky, Gluck Equine Research Center, Lexington, KY 40546-0099, USA |
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Abstract: | A novel gene, BgP12, encoding a 12-kDa protein was identified from Babesia gibsoni. The full-length cDNA of BgP12 contains an open reading frame of 378 bp, corresponding to 126 amino acid (aa) residues consisting of a putative 26 aa signal peptide and a 100 aa mature protein. The recombinant BgP12 (rBgP12) lacking the N-terminal signal peptide was expressed in Escherichia coli as a soluble glutathione S-transferase (GST) fusion protein (rBgP12) that produced an anti-rBgP12 serum in mice after immunization. Using this anti-rBgP12 serum, a native 12-kDa protein in B. gibsoni was recognized by Western blot analysis. Immunofluorescent antibody tests (IFAT) revealed that BgP12 was mainly seen during the ring stage of B. gibsoni trophozoite. An indirect enzyme-linked immunosorbent assay (ELISA) using the rBgP12 detected specific antibodies in the sequential sera of a dog experimentally infected with B. gibsoni beginning 10 days post-infection to 442 days post-infection, even when the dog became chronically infected and showed a low level of parasitemia. Moreover, the antigen did not show cross-reaction with antibodies to the closely related apicomplexan parasites, indicating that the rBgP12 might be an immunodominant antigen for B. gibsoni infection that could be used as a diagnostic antigen for B. gibsoni infection with high specificity and sensitivity. |
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