Molecular Cloning of a
            <i>Babesia caballi</i>
            Gene Encoding the 134-Kilodalton Protein and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

Tamaki, Yoh; Hirata, Haruyuki; Takabatake, Noriyuki; Bork, Sabine; Yokoyama, Naoki; Xu, Xuan; Fujisaki, Kozo; Igarashi, Ikuo

doi:10.1128/cdli.11.1.211-215.2004

Cited by 5 publications

(6 citation statements)

References 21 publications

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“…The MAEBL is an erythrocyte-binding protein located in the rhoptries and on the surface of mature merozoites; it is expressed at the beginning of schizogony (3,18). P200 was previously identified as a diagnostic antigen for the serological detection of B. bigemina infection and also has a glutamic acid-rich region, as does the Be158 protein (23). Taken together, these findings indicate that the Be158 gene product might be a novel candidate for a vaccine molecule as well as a diagnostic antigen for B. equi infection.…”

Section: Resultsmentioning

confidence: 99%

“…The resulting plasmid, designated pGEX/Be158, was used to transform the E. coli BL21 strain (Stratagene, La Jolla, Calif.) and express the recombinant Be158 gene product fused with glutathione S-transferase (GST), designated GST/Be158 protein, by standard techniques (21). The GST/Be158 protein was purified from the soluble fraction with glutathione-Sepharose 4B (Amersham Pharmacia Biotech), as described previously (9,14,23).…”

Section: Methodsmentioning

confidence: 99%

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Cloning of a Novel Babesia equi Gene Encoding a 158-Kilodalton Protein Useful for Serological Diagnosis

Hirata

Yokoyama

et al. 2005

Clin Vaccine Immunol

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In this study, we characterized a Babesia equi Be158 gene obtained by immunoscreening a B. equi cDNA expression phage library with B. equi-infected horse serum. The Be158 gene consists of an open reading frame of 3,510 nucleotides. The recombinant Be158 gene product was produced in Escherichia coli and used for the immunization of mice. In Western blot analysis, mouse immune serum against the Be158 gene product recognized 75-and 158-kDa proteins from the lysate of B. equi-infected erythrocytes. In an indirect fluorescentantibody test with the mouse immune serum, the Be158 antigen appeared in the cytoplasm of Maltese cross-forming parasites (which consist of four merozoites) and was located mainly in the extraerythrocytic merozoite body. When the recombinant Be158 gene product was used in an enzyme-linked immunosorbent assay as a serological antigen, it was found to react to B. equi-infected horse sera, indicating that the Be158 gene product is useful as a serologically diagnostic antigen for B. equi infection.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Cloning of a Novel Babesia equi Gene Encoding a 158-Kilodalton Protein Useful for Serological Diagnosis

Hirata

Yokoyama

et al. 2005

Clin Vaccine Immunol

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“…Currently, recombinant proteins are used to detect the infection rather than using a serological test with a crude antigen prepared from merozoites [7,18]. A previous study by Ikadai et al, [22] concluded that the 48-kDa protein is immunodominant and might be an important antigen for diagnosis and was reported to be involved in the invasion of erythrocytes [13] and to differentiate very clearly between B. caballi-infected horse sera, and T. equi-infected horse sera. Moreover, an ELISA using a newly identified rBC134 protein was developed and proved to be highly specific for the detection of B. caballi-specific equine antibodies [23].…”

Section: Plos Onementioning

confidence: 99%

“…Bc48 is one rhoptry protein of merozoites of B. caballi, which was previously evaluated as a promising antigen for the serological detection of antibodies of B. caballi [12]. Moreover, BC134 protein was developed and proved to be highly specific for the detection of B. caballispecific equine antibodies [13].…”

Section: Introductionmentioning

confidence: 99%

Development of a promising antigenic cocktail for the global detection of Babesia caballi in horse by ELISA

et al. 2023

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In this study, we designed novel truncated Babesia caballi (B. caballi) recombinant proteins from the previously used B. caballi proteins; 134-Kilodalton Protein (rBC134) and Merozoite Rhoptry 48 Protein (rBC48). Then, we evaluated the diagnostic performance of the newly designed proteins when used as a single antigen or when used as cocktail antigen consists of rBC134 full length (rBC134f) + newly designed rBC48 (rBC48t) or newly designed rBC134 (rBC134t) + rBC48t for the detection of B. caballi infection in horse using indirect enzyme-linked immunosorbent assay (iELISA). We used one dose and a half of each antigen in the cocktail formulas. The serum samples were collected from different endemic areas in addition to the sera collected from horses experimentally infected with B. caballi were used in the present study. Cocktail antigen in full dose of (rBC134f + rBC48t) exhibited the highest optical density (OD) values with B. caballi–infected sera and showed the lowest OD values with normal equine sera or B. caballi, and Theileria equi mixed infected sera in comparison with the single antigen. Interestingly, the same cocktail antigen exhibited the highest concordance rate (76.74%) and kappa value (0.79) in the screening of 200 field serum samples collected from five B. caballi endemic countries, including South Africa (n = 40), Ghana (n = 40), Mongolia (n = 40), Thailand (n = 40), and China (n = 40) using iELISA and the results were compared to those of indirect fluorescent antibody test (IFAT) as a reference. Moreover, the identified promising cocktail full dose antigen (rBC134f + rBC48t) showed that it can detect the infection as early as the 4th day post-infection in sera collected from experimentally infected horses. The obtained results revealed the reliability of the rBC134f + rBC48t cocktail antigen when used in full dose for the detection of specific antibodies to B. caballi in horses which will be useful for epidemiological surveys and control of equine babesiosis.

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“…Various genetic markers have been employed for the differentiation and characterization of B. caballi, comprising the BC48 and 134 protein genes (Tomaki et al, 2004), the rhoptryassociated protein (RAP-1) gene , the 16 S rRNA gene and 18S rRNA gene . Of all these genes, the 18S rRNA gene remained the most common target gene in various investigations.…”

Section: In Babesia Caballi Speciesmentioning

confidence: 99%

Equine piroplasmosis in Greece

Kenmogne¹

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Molecular Cloning of a Babesia caballi Gene Encoding the 134-Kilodalton Protein and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

Cited by 5 publications

References 21 publications

Cloning of a Novel Babesia equi Gene Encoding a 158-Kilodalton Protein Useful for Serological Diagnosis

Cloning of a Novel Babesia equi Gene Encoding a 158-Kilodalton Protein Useful for Serological Diagnosis

Development of a promising antigenic cocktail for the global detection of Babesia caballi in horse by ELISA

Equine piroplasmosis in Greece

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