Evaluation of Babesia bigemina 200 kDa recombinant antigen in enzyme-linked immunosorbent assay.
A truncated fragment of the gene encoding the 200-kDa protein (P200) of Babesia bigemina was cloned into a plasmid vector, pGEX-4 T-1 and expressed in Escherichia coli as a glutathione-S-transferase fused protein. An indirect enzyme-linked immunosorbent assay (ELISA) using the rp200/CT detected specific antibodies in cattle experimentally infected with B. bigemina. Furthermore, the antigen did not cross-react with antibodies to Babesia bovis, a closely related Babesia parasite indicating that rp200/CT is a specific antigen for the diagnosis of B. bigemina infection. Additionally, ELISA using p200/CT and polymerase chain reaction were conducted on serum and corresponding DNA samples obtained from field cattle to evaluate the diagnostic utility of the p200/CT antigen. Results from the current study suggest that p200/CT ELISA is a sensitive and specific method for improved serodiagnosis of B. bigemina infection.
Altangerel
Khukhuu
K
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
Alhassan
Andy
A
Iseki
Hiroshi
H
Sivakumar
Thillaiampalam
T
Boldbaatar
Damdinsuren
D
Yokoyama
Naoaki
N
Igarashi
Ikuo
I
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000235:genetics; D001403:Babesia / Q000737:chemistry* / Q000235:genetics; D001404:Babesiosis / Q000175:diagnosis / Q000662:veterinary*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D003001:Cloning, Molecular; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012680:Sensitivity and Specificity
C411107:200kDa Babesia bigemina antigen; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D011993:Recombinant Fusion Proteins