Development of an ELISA using anti-idiotypic antibody for diagnosis of opisthorchiasis
Language English Country Czech Republic Media electronic
Document type Journal Article
PubMed
27507639
DOI
10.14411/fp.2016.025
PII: 2016.025
Knihovny.cz E-resources
- Keywords
- Opisthorchis felineus, antigenicity, excretory-secretory antigen, immunogenicity, monoclonal antibody, polyclonal antibody,
- MeSH
- Enzyme-Linked Immunosorbent Assay * MeSH
- Humans MeSH
- Opisthorchis MeSH
- Opisthorchiasis blood diagnosis MeSH
- Antibodies, Anti-Idiotypic metabolism MeSH
- Sensitivity and Specificity MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Antibodies, Anti-Idiotypic MeSH
Monoclonal antibody specific for an epitope of cretory-secretory antigen protein of Opisthorchis felineus (Rivolta, 1884) (Trematoda: Opisthorchiidae) with a molecular weight of 28 kDa was used in a sandwich enzyme-linked immunosorbent assay (ELISA) for immobilisation of liver fluke specific antigen to the solid phase. Examination of human sera by this ELISA compared with commercial assays demonstrated that the monoclonal antibody epitope is located within this significant parasite protein. Anti-idiotypic antibody specific for the paratope of this monoclonal antibody was obtained by a hybridoma technique. Mimicking an epitope of excretory-secretory antigen of O. felineus, it had the capacity to bind specific antibody and elicit an antibody response. The value of anti-idiotypic antibody as a substitute for the liver fluke antigen was tested by ELISA using serum samples of infected dogs. Anti-idiotypic antibody proved to be of value in both an indirect-ELISA and a competitive-ELISA for diagnosis of opisthorchiasis. Mature trematodes were isolated from all infected animals. The faecal egg counts were negative in dogs with a relatively small number of parasites, despite finding antibodies in serum by ELISA. Substitution of parasite antigen with anti-idiotype avoids the use of experimental animals and also reduces time-consuming steps of antigen preparation.
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