Previous Article | Next Article 
Journal of Clinical Microbiology, November 2001, p. 3987-3991, Vol. 39, No. 11
National Research Center for Protozoan
Diseases, Obihiro University of Agriculture and Veterinary Medicine,
Obihiro, Hokkaido 080-8555,1 and College
of Bioresource Science, Nihon University, Fujisawa, Kanagawa
252-8510,3 Japan, and Department of
Clinical Microbiology, Faculty of Associated Medical Sciences,
Chiang Mai University, Chiang Mai 50200, Thailand2
Received 10 May 2001/Returned for modification 10 July
2001/Accepted 12 August 2001
The baculovirus expression system has proved to be a
useful tool for the production of recombinant proteins. Here
we have characterized the Neospora caninum surface
protein NcSRS2 produced by two types of the recombinant
virus and also have developed an enzyme-linked immunosorbent assay
(ELISA) using recombinant NcSRS2 for the serologic diagnosis of
Neospora infection. Western blot analysis showed two
major protein bands that were detectable in insect cells infected with
each recombinant baculovirus, and a lower-molecular-weight protein
was detected in culture supernatants from a cell infected with the
recombinant virus lacking the hydrophobic C-terminal tail. Analysis of
the N-terminal amino acids showed that the secreted NcSRS2 lacked 6 kDa
of the N-terminal signal peptide. Moreover, the detergent-soluble
protein of insect cells infected with the recombinant baculovirus
expressing the full-length NcSRS2 gene was used to develop an ELISA
system based on specificity and reactivity to antisera against
Toxoplasma gondii, Hammondia heydorni, or
N. caninum. Anti-N.
caninum mouse, dog, and bovine sera recognized
the recombinant NcSRS2 on Western blots. Furthermore, we have shown
that the developed ELISA system consistently discriminates indirect
fluorescent-antibody test (IFAT)-positive bovine sera against N.
caninum from IFAT-negative sera. These results indicate that
the ELISA using baculovirus-expressed NcSRS2 can be useful for
effective and reliable serodiagnosis of N. caninum infection.
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.11.3987-3991.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Characterization of Neospora caninum Surface
Protein NcSRS2 Based on Baculovirus Expression System and Its
Application for Serodiagnosis of Neospora
Infection
*
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: 81-155-49-5644. Fax:
81-155-49-5643. E-mail: nrcpmi{at}obihiro.ac.jp.
Journal of Clinical Microbiology, November 2001, p. 3987-3991, Vol. 39, No. 11
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.11.3987-3991.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Ghalmi, F., China, B., Kaidi, R., Losson, B.
(2009). Evaluation of a SRS2 sandwich commercial enzyme-linked immunosorbent assay for the detection of anti-Neospora caninum antibodies in bovine and canine sera. jvdi
21: 108-111
[Abstract] [Full Text] -
Hoane, J. S., Morrow, J. K., Saville, W. J., Dubey, J. P., Granstrom, D. E., Howe, D. K.
(2005). Enzyme-Linked Immunosorbent Assays for Detection of Equine Antibodies Specific to Sarcocystis neurona Surface Antigens. CVI
12: 1050-1056
[Abstract] [Full Text] -
Liao, M., Zhang, S., Xuan, X., Zhang, G., Huang, X., Igarashi, I., Fujisaki, K.
(2005). Development of Rapid Immunochromatographic Test with Recombinant NcSAG1 for Detection of Antibodies to Neospora caninum in Cattle. CVI
12: 885-887
[Abstract] [Full Text]
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»