Evaluation of Babesia bigemina 200 kDa recombinant antigen in enzyme-linked immunosorbent assay
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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.
KeywordsPolymerase Chain Reaction Babesia Babesiosis Infected Cattle Ampicillin Sodium
This study was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science, a Program for Promotion of Basic Research Activities for Innovative Biosciences, and grants from the twenty-first century COE Program and the Program of Founding Research Centers for Emerging and Reemerging Infectious Diseases, the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and the Japan International Cooperation Agency.
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