Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide
Brucella melitensis native haptens (NH) are polysaccharides identical to the O-side chain of the smooth lipopolysaccharide (S-LPS) (E. Moreno, H. Mayer, and I. Moriyón, Infect. Immun. 55:2850-2853, 1987) which precipitate with sera from infected cattle but not from strain 19-vaccinated cattle. In th...
| Main Authors: | , , , |
|---|---|
| Format: | info:eu-repo/semantics/article |
| Language: | eng |
| Published: |
American Society for Microbiology
2013
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10171/29426 |
| _version_ | 1793400171173773312 |
|---|---|
| author | Alonso-Urmeneta, B. (Begoña) Moriyon, I. (Ignacio) Diaz, R. (Ramón) Blasco, J.M. (J. M.) |
| author_facet | Alonso-Urmeneta, B. (Begoña) Moriyon, I. (Ignacio) Diaz, R. (Ramón) Blasco, J.M. (J. M.) |
| author_sort | Alonso-Urmeneta, B. (Begoña) |
| collection | DSpace |
| description | Brucella melitensis native haptens (NH) are polysaccharides identical to the O-side chain of the smooth lipopolysaccharide (S-LPS) (E. Moreno, H. Mayer, and I. Moriyón, Infect. Immun. 55:2850-2853, 1987) which precipitate with sera from infected cattle but not from strain 19-vaccinated cattle. In the present work, NH was extracted by the hot-water method (R. Díaz, J. Toyos, M.D. Salvo, and M.L. Pardo, Ann. Rech. Vet. 12:35-39, 1981) and purified free of S-LPS and protein. Purified NH lacked the ability to coat polystyrene and sheep erythrocytes. In contrast, NH acylated with stearoyl chloride bound to both polystyrene and erythrocytes. By hemagglutination and enzyme-linked immunosorbent assay (ELISA), S-LPS and acylated NH gave similar results with blood sera from brucellosis-free, strain 19-vaccinated, and infected cattle. Moreover, a significant correlation between the results of NH ELISA and S-LPS ELISA was demonstrated with milk sera. However, in a competitive ELISA with milk sera, S-LPS in the liquid phase abrogated the binding of antibodies to acylated NH adsorbed to polystyrene, while NH in the liquid phase did not influence the binding of antibodies to polystyrene-adsorbed S-LPS. It is hypothesized that the different precipitations of NH and S-LPS with sera from infected or strain 19-vaccinated cattle are due to differences in the affinity of the antibodies produced upon vaccination or infection and in the physical state of aggregation of NH and S-LPS in aqueous solutions. |
| format | info:eu-repo/semantics/article |
| id | oai:dadun.unav.edu:10171-29426 |
| institution | Universidad de Navarra |
| language | eng |
| publishDate | 2013 |
| publisher | American Society for Microbiology |
| record_format | dspace |
| spelling | oai:dadun.unav.edu:10171-294262020-03-03T23:37:38Z Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide Alonso-Urmeneta, B. (Begoña) Moriyon, I. (Ignacio) Diaz, R. (Ramón) Blasco, J.M. (J. M.) Brucella immunology Haptens analysis Lipopolysaccharides analysis Polysaccharides Bacterial analysis Brucella melitensis native haptens (NH) are polysaccharides identical to the O-side chain of the smooth lipopolysaccharide (S-LPS) (E. Moreno, H. Mayer, and I. Moriyón, Infect. Immun. 55:2850-2853, 1987) which precipitate with sera from infected cattle but not from strain 19-vaccinated cattle. In the present work, NH was extracted by the hot-water method (R. Díaz, J. Toyos, M.D. Salvo, and M.L. Pardo, Ann. Rech. Vet. 12:35-39, 1981) and purified free of S-LPS and protein. Purified NH lacked the ability to coat polystyrene and sheep erythrocytes. In contrast, NH acylated with stearoyl chloride bound to both polystyrene and erythrocytes. By hemagglutination and enzyme-linked immunosorbent assay (ELISA), S-LPS and acylated NH gave similar results with blood sera from brucellosis-free, strain 19-vaccinated, and infected cattle. Moreover, a significant correlation between the results of NH ELISA and S-LPS ELISA was demonstrated with milk sera. However, in a competitive ELISA with milk sera, S-LPS in the liquid phase abrogated the binding of antibodies to acylated NH adsorbed to polystyrene, while NH in the liquid phase did not influence the binding of antibodies to polystyrene-adsorbed S-LPS. It is hypothesized that the different precipitations of NH and S-LPS with sera from infected or strain 19-vaccinated cattle are due to differences in the affinity of the antibodies produced upon vaccination or infection and in the physical state of aggregation of NH and S-LPS in aqueous solutions. 2013-06-25T07:50:06Z 2013-06-25T07:50:06Z 1988 info:eu-repo/semantics/article https://hdl.handle.net/10171/29426 eng info:eu-repo/semantics/openAccess application/pdf American Society for Microbiology |
| spellingShingle | Brucella immunology Haptens analysis Lipopolysaccharides analysis Polysaccharides Bacterial analysis Alonso-Urmeneta, B. (Begoña) Moriyon, I. (Ignacio) Diaz, R. (Ramón) Blasco, J.M. (J. M.) Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide |
| title | Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide |
| title_full | Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide |
| title_fullStr | Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide |
| title_full_unstemmed | Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide |
| title_short | Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide |
| title_sort | enzyme-linked immunosorbent assay with brucella native hapten polysaccharide and smooth lipopolysaccharide |
| topic | Brucella immunology Haptens analysis Lipopolysaccharides analysis Polysaccharides Bacterial analysis |
| url | https://hdl.handle.net/10171/29426 |
| work_keys_str_mv | AT alonsourmenetabbegona enzymelinkedimmunosorbentassaywithbrucellanativehaptenpolysaccharideandsmoothlipopolysaccharide AT moriyoniignacio enzymelinkedimmunosorbentassaywithbrucellanativehaptenpolysaccharideandsmoothlipopolysaccharide AT diazrramon enzymelinkedimmunosorbentassaywithbrucellanativehaptenpolysaccharideandsmoothlipopolysaccharide AT blascojmjm enzymelinkedimmunosorbentassaywithbrucellanativehaptenpolysaccharideandsmoothlipopolysaccharide |