Serodiagnosis of canine Babesia gibsoni infection by enzyme-linked immunosorbent assay with recombinant P50 expressed in Escherichia coli.
The entire P50 gene encoding a surface protein of Babesia gibsoni was cloned into the bacteria expression vector pGEX-4T-3 and subsequently expressed in Escherichia coli as a glutathione S-transferase fusion protein. The purified recombinant P50 was evaluated in an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of B. gibsoni infection in dogs. ELISA was able to differentiate clearly among B. gibsoni-infected, Babesia canis-infected, and uninfected dog sera. The antibody response against the recombinant P50 was maintained at a high level until the chronic stage of infection in dogs experimentally infected with B. gibsoni. When serum samples collected from domestic dogs in Japan were examined for the diagnosis of B. gibsoni infection by the ELISA, 3 of 209 samples (1.4%) were positive for the antibody to B. gibsoni. This result was completely identical to those of Western blot analysis and the indirect fluorescent antibody test. These results indicate that the recombinant P50 expressed in E. coil is a useful diagnostic antigen for practical use in the diagnosis of B. gibsoni infection in dogs.
Fukumoto
Shinya
S
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Sekine
Yukiko
Y
Xuan
Xuenan
X
Igarashi
Ikuo
I
Sugimoto
Chihiro
C
Nagasawa
Hideyuki
H
Fujisaki
Kozo
K
Mikami
Takeshi
T
Suzuki
Hiroshi
H
D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000097:blood; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology; D001404:Babesiosis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D015153:Blotting, Western / Q000662:veterinary; D004283:Dog Diseases / Q000175:diagnosis* / Q000276:immunology; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D005786:Gene Expression Regulation; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology; D051379:Mice; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D016056:Immunodominant Epitopes; C434466:P50 protein, Babesia gibsoni; D011994:Recombinant Proteins