dc.identifier.citation | Mousa A.A, Cao S, Aboge G.O, Terkawi MA, El Kirdasy A, Salama A, Attia M, Aboulaila M, Zhou M, Kamyingkird K, Moumouni PF, Masatani T, El Aziz SA, Moussa WM, Chahan B, Fukumoto S, Nishikawa Y, El Ballal SS, Xuan X.(2013).ExpParasitol. 2013 Oct; 135(2):414-20 | en_US |
dc.description.abstract | Identification and molecular characterization of Babesia gibsoni proteins with
potential antigenic properties are crucial for the development and validation of the
serodiagnostic method. In this study, we isolated a cDNA clone encoding a novel B. gibsoni
76-kDa protein by immunoscreening of the parasite cDNA library. Computer analysis
revealed that the protein presents a glutamic acid-rich region in the C-terrninal. Therefore, the
protein was designated as B. gibsoni glutamic acid-rich protein (BgGARP). A Blastp analysis
of a translated BgGARP polypeptide demonstrated that the peptide shared a significant
homology with a 200-kDa protein of Babesiabigemina and Babesiabovis. A truncated
BgGARP cDNA (BgGARPt) encoding a predicted 13-kDa peptide was expressed in
Escherichia coli (E. coli), and mouse antisera against the recombinant protein were used to
characterize a corresponding native protein. The antiserum against recombinant BgGARPt
(rBgGARPt) recognized a 140-kDa protein in the lysate of infected erythrocytes, which was
detectable in the cytoplasm of the parasites by confocal microscopic observation. In addition,
the specificity and sensitivity of enzyme-linked immunosorbent assay (ELISA) with
rBgGARPt were evaluated using B. gibsoni-infected dog sera and specific pathogen-free
(SPF) dog sera. Moreover, 107 serum samples from dogs clinically diagnosed with babesiosis
were examined using ELISA with rBgGARPt. The results showed that 86 (80.4%) samples
were positive by rBgGARPt-ELISA, which was comparable to IFAT and PCR as reference
test. Taken together, these results demonstrate that BgGARP is a suitable serodiagnostic
antigen for detecting antibodies against B. gibsoni in dogs. | en_US |