Detection of antibodies to Neospora caninum in cattle by enzyme-linked immunosorbent assay with truncated NcSRS2 expressed in Escherichia coli.
The surface antigen 1-related sequence 2 of Neospora caninum (NcSRS2) is considered as an immunodominant antigen. In this study, the gene encoding truncated NcSRS2 (NcSRS2t) lacking an N-terminal signal peptide and C-terminal hydrophobic regions was expressed in Escherichia coli, and its diagnostic potential in an enzyme-linked immunosorbent assay (ELISA) was evaluated. ELISA could discriminate clearly between known N. caninum-positive and -negative sera from cattle. Field serum samples collected from cattle in Brazil were examined for the diagnosis of N. caninum infection using ELISA. Of the 197 samples analyzed, 64 (32.5%) samples were positive for antibodies to N. caninum. Of the 64 ELISA-positive samples, 58 (90.6%) were confirmed as positive by Western blot analysis with whole-parasite antigens. These results suggest that ELISA with recombinant NcSRS2t is an effective method for diagnosis of N. caninum infection in cattle.
Gaturaga
Irungu
I
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. gen@obihiro.ac.jp
Chahan
Bayin
B
Xuan
Xuenan
X
Huang
Xiaohong
X
Liao
Min
M
Fukumoto
Shinya
S
Hirata
Haruyuki
H
Nishikawa
Yoshihumi
Y
Takashima
Yasuhiro
Y
Suzuki
Hiroshi
H
Fujisaki
Kozo
K
Sugimoto
Chihiro
C
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology*; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western / Q000662:veterinary; D001938:Brazil / Q000453:epidemiology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000453:epidemiology / Q000469:parasitology; D003048:Coccidiosis / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary*; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D051379:Mice; D008807:Mice, Inbred BALB C; D018688:Neospora / Q000276:immunology*; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D016036:Seroepidemiologic Studies
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C115389:SRS2 protein, Neospora caninum