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Journal of Clinical Microbiology
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Clinical Veterinary Microbiology

Expression of Babesia equi Merozoite Antigen 1 in Insect Cells by Recombinant Baculovirus and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

Xuenan Xuan, Alejandra Larsen, Hiromi Ikadai, Tetsuya Tanaka, Ikuo Igarashi, Hideyuki Nagasawa, Kozo Fujisaki, Yutaka Toyoda, Naoyoshi Suzuki, Takeshi Mikami
Xuenan Xuan
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Alejandra Larsen
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Hiromi Ikadai
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Tetsuya Tanaka
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Ikuo Igarashi
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Hideyuki Nagasawa
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Kozo Fujisaki
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Yutaka Toyoda
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Naoyoshi Suzuki
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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Takeshi Mikami
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
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DOI: 10.1128/JCM.39.2.705-709.2001
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ABSTRACT

The gene encoding the entire Babesia equi merozoite antigen 1 (EMA-1) was inserted into a baculovirus transfer vector, and a recombinant virus expressing EMA-1 was isolated. The expressed EMA-1 was transported to the surface of infected insect cells, as judged by an indirect fluorescent-antibody test (IFAT). The expressed EMA-1 was also secreted into the supernatant of a cell culture infected with recombinant baculovirus. Both intracellular and extracellular EMA-1 reacted with a specific antibody in Western blots. The expressed EMA-1 had an apparent molecular mass of 34 kDa that was identical to that of native EMA-1. The secreted EMA-1 was used as an antigen in an enzyme-linked immunosorbent assay (ELISA). The ELISA differentiatedB. equi-infected horse sera from Babesia caballi-infected horse sera or normal horse sera. The ELISA was more sensitive than the complement fixation test and IFAT. These results demonstrated that the recombinant EMA-1 expressed in insect cells might be a useful diagnostic reagent for detection of antibodies to B. equi.

  • Copyright © 2001 American Society for Microbiology
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Expression of Babesia equi Merozoite Antigen 1 in Insect Cells by Recombinant Baculovirus and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay
Xuenan Xuan, Alejandra Larsen, Hiromi Ikadai, Tetsuya Tanaka, Ikuo Igarashi, Hideyuki Nagasawa, Kozo Fujisaki, Yutaka Toyoda, Naoyoshi Suzuki, Takeshi Mikami
Journal of Clinical Microbiology Feb 2001, 39 (2) 705-709; DOI: 10.1128/JCM.39.2.705-709.2001

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Expression of Babesia equi Merozoite Antigen 1 in Insect Cells by Recombinant Baculovirus and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay
Xuenan Xuan, Alejandra Larsen, Hiromi Ikadai, Tetsuya Tanaka, Ikuo Igarashi, Hideyuki Nagasawa, Kozo Fujisaki, Yutaka Toyoda, Naoyoshi Suzuki, Takeshi Mikami
Journal of Clinical Microbiology Feb 2001, 39 (2) 705-709; DOI: 10.1128/JCM.39.2.705-709.2001
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KEYWORDS

Babesia
babesiosis
Horse Diseases
Membrane Proteins
Protozoan Proteins

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