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Your search keyword '"Goo, Y.-K."' showing total 2 results
Start Over Author "Goo, Y.-K." Topic enzyme-linked immunosorbent assay
2 results on '"Goo, Y.-K."'

1. Identification of a novel B. gibsoni 27-kDa protein as a serodiagnostic antigen

Author

Terkawi, M. A., Aboge, G., Jia, H., Goo, Y-K., Ooka, H., Yamagishi, Junya, Nishikawa, Yoshifumi, Kawazu, Shin-ichiro, Fujisaki, K., and Xuan, Xuenan

Subjects
Babesia gibsoni, deiagnostic performance, enzyme-linked immunosorbent assay
Abstract

application/pdf, A novel gene encoding 27-kDa protein was identified by the screening of Babesia gibsoni cDNA library with acutely infected dog serum. The BgP27 is a single copy gene with a predicted open reading frame of 762 bp and 254 amino acids. The phylogenic analysis of the deduced amino acid of BgP27 demonstrated considerable identities with members of Plasmodium berghei circumsporozoite protein family that ranged between 18.4% and 22.8%. The BgP27 was expressed as a glutathione S-transferase fusion protein in Escherichia coli. The serum raised in mice against the recombinant protein specifically reacted with a 27-kDa protein in the extracts of B. gibsoni parasites. Confocal laser scanning microscopic observation showed high fluorescent reactivity with both extracellular and intracellular merozoite. Furthermore, recombinant BgP27 was used as an antigen in an enzyme-linked immunosorbent assay (ELISA). Thus, the kinetics of the anti-BgP27 antibody was detected in serial serum samples collected from a B. gibsoni-infected dog. IgG levels were high throughout the course of infection. In addition, the ELISA was able to differentiate between B. gibsoni-infected dog serum and B. canis subspecies-infected dog serum or normal dog serum. The diagnostic performance of BgP27-ELISA revealed the potential use of the antigen for detection of infection in dogs.

Published
2008

2. Molecular characterizations of three distinct Babesia gibsoni rhoptry-associated protein-1s (RAP-1s).

Author

Terkawi, M. A., Amornthep, A., Ooka, H., Aboge, G., Jia, H., Goo, Y.-K., Nelson, B., Yamagishi, J., Nishikawa, Y., Igarashi, I., Kawazu, S.-I., Fujisaki, K., and Xuan, X.

Subjects
RECOMBINANT proteins, PERFORMANCE-enhancing drugs, ENZYME-linked immunosorbent assay, RECOMBINANT molecules, IMMUNOLOGY technique, IMMUNOFLUORESCENCE
Abstract

Three cDNAs encoding rhoptry-associated protein 1 (RAP-1) homologues were found in the Babesia gibsoni EST database. Based on similarities to BgRAP-1a, which was identified previously by serological screening of a cDNA merozoite library, the two new genes were designated BgRAP-1b (33.7%) and BgRAP-1c (57%). Mice antiserum raised against each recombinant protein reacted specifically with B. gibsoni parasites as determined by Western blotting, which showed native molecular sizes of the BgRAP-1a (51 kDa), BgRAP-1b (53 kDa) and BgRAP-1c (47 kDa) consistent with predictable molecular weights. Immunofluoresence using these antibodies revealed localization of all BgRAP-1s within the matrix of merozoites; however, BgRAP-1a appeared to diverge from the other two when it was found secreted into the cytoplasm of infected erythrocytes. Apical localization of all 3 BgRAP-1s during the extracellular stage of the parasite combined with their ability to bind a canine erythrocyte membrane fraction was suggestive of a role for these proteins in erythrocyte attachment. Lastly, the ability of these recombinant proteins to be used as diagnostic reagents was tested by ELISA and the sensitivities of BgRAP-1a and BgRAP-1c were found increased through N-terminal truncation. Taken together, our data suggest divergent roles for the 3 BgRAP-1s in the merozoite stage of B. gibsoni. [ABSTRACT FROM AUTHOR]

Published
2009
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