title: Expression of truncated Babesia gibsoni thrombospondin-related adhesive proteins in Escherichia coli and evaluation of their diagnostic potential by enzyme-linked immunosorbent assay.
abstract: Among the previously established enzyme-linked immunosorbent assays (ELISAs), an ELISA using the full length of a recombinant thrombospondin-related adhesive protein of Babesia gibsoni (rBgTRAPf) is considered as the most sensitive diagnostic method for the detection of an antibody to B. gibsoni in dogs. However, the expression of rBgTRAPf in high concentration is poor and, thus, limits its usefulness as a diagnostic antigen. To improve its expression level, we have truncated BgTRAPf into two fragments having either an N- or a C-terminus (BgTRAPn or BgTRAPc, respectively). The expression of BgTRAPc protein in Escherichia coli yielded adequate recombinant protein. The specificity and sensitivity of ELISAs with the truncated proteins were determined using dog sera experimentally infected with B. gibsoni and specific pathogen-free (SPF) dog sera. A total of 254 field dog sera were examined by the ELISA with rBgTRAPn, rBgTRAPc, and rBgTRAPf as well as by an indirect fluorescent antibody test (IFAT). The specificity of rBgTRAPc was the highest (97.15%), and its kappa value was more (0.8003) than rBgTRAPn (0.7083). With a sufficient level of expression as well as higher specificity and reliable sensitivity, rBgTRAPc appears to be a potential candidate antigen for the serodiagnosis of B. gibsoni infection in dogs.
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Narantsatsral
authors.forename: Youn-Kyoung
authors.initials: YK
authors.lastname: Goo
authors.forename: B
authors.initials: B
authors.lastname: Battsetseg
authors.forename: P
authors.initials: P
authors.lastname: Myagmarsuren
authors.forename: Mohamad Alaa
authors.initials: MA
authors.lastname: Terkawi
authors.forename: Takehisa
authors.initials: T
authors.lastname: Soma
authors.forename: Yuzi
authors.initials: Y
authors.lastname: Luo
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Shinuo
authors.initials: S
authors.lastname: Cao
authors.forename: Longzheng
authors.initials: L
authors.lastname: Yu
authors.forename: Ketsarin
authors.initials: K
authors.lastname: Kamyingkird
authors.forename: Gabriel Oluga
authors.initials: GO
authors.lastname: Aboge
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Nishikawa
authors.forename: Xuenan
authors.initials: X
authors.lastname: Xuan
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001403:Babesia / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001404:Babesiosis / Q000175:diagnosis / Q000662:veterinary*; D015153:Blotting, Western; D004283:Dog Diseases / Q000175:diagnosis*; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D004926:Escherichia coli / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C069428:thrombospondin-related adhesive protein, protozoan