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Journal of Clinical Microbiology, April 2004, p. 1601-1604, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1601-1604.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Improved Enzyme-Linked Immunosorbent Assay Using C-Terminal Truncated Recombinant Antigens of Babesia bovis Rhoptry-Associated Protein-1 for Detection of Specific Antibodies

Suthisak Boonchit,1 Xuenan Xuan,1 Naoaki Yokoyama,1 Will L. Goff,2 Suryakant D. Waghela,3 Gale Wagner,3 and Ikuo Igarashi1*

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan,1 Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164,2 Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, Texas 778433

Received 14 April 2003/ Returned for modification 26 May 2003/ Accepted 8 August 2003

An enzyme-linked immunosorbent assay (ELISA) based on a recombinant rhoptry-associated protein-1 (RAP-1) of Babesia bovis has been previously developed, but it was imperfect because some cross-reactions were still present in Babesia bigemina-infected bovine sera. To improve its accuracy for the specific detection of the antibodies to B. bovis, we constructed three C-terminal truncated recombinant antigens of the RAP-1—rCT1 (amino acids [aa] 301 to 408), rCT2 (aa 388 to 490), and rCT3 (aa 466 to 565)—by using a baculovirus expression system and evaluated their diagnostic potentials using ELISA. rCT1 and rCT2 were better diagnostic antigens in their sensitivities and diagnostic efficiencies than rCT3, although none of the recombinant antigens showed any cross-reactivity to B. bigemina-infected bovine sera. These results confirmed that the N-terminal 300-aa region caused cross-reactivity of the entire RAP-1 antigen, and the C-terminal truncated recombinant antigens were shown to be useful reagents for species-specific serodiagnosis.

* Corresponding author. Mailing address: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. Phone: 81-155-49-5641. Fax: 81-155-49-5643. E-mail: igarcpmi{at}obihiro.ac.jp.

Journal of Clinical Microbiology, April 2004, p. 1601-1604, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1601-1604.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kim, C.-M., Blanco, L. B. C., Alhassan, A., Iseki, H., Yokoyama, N., Xuan, X., Igarashi, I. (2008). Development of a Rapid Immunochromatographic Test for Simultaneous Serodiagnosis of Bovine Babesioses Caused by Babesia bovis and Babesia bigemina. Am J Trop Med Hyg 78: 117-121 [Abstract] [Full Text]  


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