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Journal of Clinical Microbiology, October 2002, p. 3771-3775, Vol. 40, No. 10
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.10.3771-3775.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Evaluation of an Enzyme-Linked Immunosorbent Assay with Recombinant Rhoptry-Associated Protein 1 Antigen against Babesia bovis for the Detection of Specific Antibodies in Cattle

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

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan,1 Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630,2 Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, Texas 778433

Received 6 May 2002/ Returned for modification 17 June 2002/ Accepted 8 July 2002

The gene encoding Babesia bovis rhoptry-associated protein 1 (RAP-1) was used to develop an enzyme-linked immunosorbent assay (ELISA) to measure specific antibodies against B. bovis. The B. bovis RAP-1 gene was subcloned into a baculovirus transfer vector, and the RAP-1 protein was expressed in insect cells infected with a recombinant baculovirus. The recombinant B. bovis RAP-1 of 65 kDa was detected with anti-RAP-1 mouse serum by Western blotting, and this recombinant RAP-1 was used as an antigen in the ELISA. The ELISA was able to differentiate between B. bovis-infected sera and B. bigemina-infected sera or noninfected normal bovine sera. The results demonstrate that the recombinant RAP-1 expressed in insect cells might be a useful antigen for the detection of antibodies to B. bovis.

* 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, October 2002, p. 3771-3775, Vol. 40, No. 10
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.10.3771-3775.2002
Copyright © 2002, 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]  
  • Boonchit, S., Xuan, X., Yokoyama, N., Goff, W. L., Waghela, S. D., Wagner, G., Igarashi, I. (2004). Improved Enzyme-Linked Immunosorbent Assay Using C-Terminal Truncated Recombinant Antigens of Babesia bovis Rhoptry-Associated Protein-1 for Detection of Specific Antibodies. J. Clin. Microbiol. 42: 1601-1604 [Abstract] [Full Text]  


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