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Journal of clinical microbiology2001; 39(2); 705-709; doi: 10.1128/JCM.39.2.705-709.2001

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.

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 differentiated B. 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.
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Publication Date: 2001-02-07 PubMed ID: 11158131PubMed Central: PMC87800DOI: 10.1128/JCM.39.2.705-709.2001Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research presents a breakthrough in the ability to differentiate antibodies between Babesia equi (a parasite affecting horses) and other types of infections. The researchers used a baculovirus, a virus commonly used in genetics research, to deliver, express, and isolate the antigen EMA-1. This expressed EMA-1 was evaluated as a diagnostic tool in enzyme-linked immunosorbent assay (ELISA) testing and displayed greater sensitivity compared with traditional diagnostic approaches.

Research Objectives and Approach

  • The research aimed to express EMA-1 (merozoite antigen 1 from Babesia equi) using a baculovirus transfer vector to isolate a recombinant virus expressing this antigen.
  • The researchers also aimed to evaluate if the expressed EMA-1 could be used as a diagnostic tool to differentiate between B. equi-infected horse serum and that of other similar infections.

Findings and Methodology

  • The research findings confirmed that the EMA-1 antigen could be expressed and transported to the surface of infected insect cells using a baculovirus transfer vector.
  • This was ascertained using an indirect fluorescent-antibody test (IFAT). Additionally, EMA-1 could also be secreted into the supernatant of a cell culture infected with the recombinant baculovirus.
  • The secreted and intracellular EMA-1 reacted with a specific antibody in Western blot tests, indicative of successful expression.
  • The researchers further found that the expressed EMA-1 matched the molecular mass of native EMA-1, providing more evidence of successful replication.

Potential Application

  • The recombinant EMA-1 expressed was used as an antigen in an ELISA, a test commonly used in medicine to detect substances that have antigenic properties, such as antibodies and peptides.
  • The ELISA was successful in differentiating between B. equi-infected horse sera from Babesia caballi-infected horse sera or normal horse sera.
  • Furthermore, this approach showed more sensitivity than the traditional diagnostic tools such as the complement fixation test and IFAT, suggesting that the recombinant EMA-1 expressed in insect cells could be a valuable diagnostic reagent for detection of antibodies to B. equi.

Cite This Article

APA
Xuan X, Larsen A, Ikadai H, Tanaka T, Igarashi I, Nagasawa H, Fujisaki K, Toyoda Y, Suzuki N, Mikami T. (2001). 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. J Clin Microbiol, 39(2), 705-709. https://doi.org/10.1128/JCM.39.2.705-709.2001

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 39
Issue: 2
Pages: 705-709

Researcher Affiliations

Xuan, X
  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Larsen, A
    Ikadai, H
      Tanaka, T
        Igarashi, I
          Nagasawa, H
            Fujisaki, K
              Toyoda, Y
                Suzuki, N
                  Mikami, T

                    MeSH Terms

                    • Animals
                    • Antibodies, Protozoan / analysis
                    • Antigens, Protozoan / analysis
                    • Antigens, Protozoan / genetics
                    • Babesia / genetics
                    • Babesia / isolation & purification
                    • Babesiosis / diagnosis
                    • Babesiosis / immunology
                    • Babesiosis / veterinary
                    • Cell Line
                    • Enzyme-Linked Immunosorbent Assay
                    • Erythrocytes / parasitology
                    • Fluorescent Antibody Technique, Indirect
                    • Genes, Protozoan
                    • Genetic Vectors
                    • Horse Diseases / diagnosis
                    • Horse Diseases / immunology
                    • Horse Diseases / parasitology
                    • Horses
                    • Membrane Proteins / analysis
                    • Membrane Proteins / genetics
                    • Mice
                    • Nucleopolyhedroviruses
                    • Polymerase Chain Reaction
                    • Protozoan Proteins / analysis
                    • Protozoan Proteins / genetics
                    • Recombinant Proteins / analysis
                    • Spodoptera

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