title: Molecular characterization and antigenic properties of a novel Babesia gibsoni glutamic acid-rich protein (BgGARP).
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-terminal. 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 Babesia bigemina and Babesia bovis. 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.
authors: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan; Department of Biochemistry and Chemistry of Nutrition, Faculty of Veterinary Medicine, Sadat City University, Sadat City, Menoufyia, Egypt.
authors: Ahmed Abdelmoniem
authors: AA
authors: Mousa
authors: Shinuo
authors: S
authors: Cao
authors: Gabriel Oluga
authors: GO
authors: Aboge
authors: Mohamad Alaa
authors: MA
authors: Terkawi
authors: Ahmed
authors: A
authors: El Kirdasy
authors: Akram
authors: A
authors: Salama
authors: Mabrouk
authors: M
authors: Attia
authors: Mahmoud
authors: M
authors: Aboulaila
authors: Mo
authors: M
authors: Zhou
authors: Ketsarin
authors: K
authors: Kamyingkird
authors: Paul Franck Adjou
authors: PF
authors: Moumouni
authors: Tatsunori
authors: T
authors: Masatani
authors: Sami Ahmed Abd
authors: SA
authors: El Aziz
authors: Waheed Mohammed
authors: WM
authors: Moussa
authors: Bayin
authors: B
authors: Chahan
authors: Shinya
authors: S
authors: Fukumoto
authors: Yoshifumi
authors: Y
authors: Nishikawa
authors: Salah Sayed
authors: SS
authors: El Ballal
authors: Xuenan
authors: X
authors: Xuan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology*; D001403:Babesia / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D001404:Babesiosis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary; D001483:Base Sequence; D003001:Cloning, Molecular; D003429:Cross Reactions; D004283:Dog Diseases / Q000175:diagnosis / Q000469:parasitology; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards / Q000662:veterinary; D005260:Female; D015870:Gene Expression; D018698:Glutamic Acid; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms
chemical_list: D000953:Antigens, Protozoan; D007106:Immune Sera; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D018698:Glutamic Acid
keywords: Babesia gibsoni; BgGARP; ELISA