Antigenic profiles of recombinant proteins from Mycobacterium avium subsp. paratuberculosis in sheep with Johne's
John P Bannantine1, Valentina Rosu, Stefania Zanetti
1National Animal Disease Center, USDA-Agricultural Research Service, Ames, IA 50010, USA.
Abstract:
Methods to improve the ELISA test to detect Mycobacterium avium subsp. paratuberculosis have been explored over several years. Previously, selected recombinant proteins of M. avium subspecies paratuberculosis were found to be immunogenic in cattle with Johne's disease. In the present study, antibody responses of infected and healthy sheep were evaluated using 18 purified recombinant proteins in an ELISA-based format for the serodiagnosis of ovine paratuberculosis. These selected recombinant proteins represent heat shock proteins, hypothetical proteins and cell surface proteins of M. avium subsp. paratuberculosis. Whereas, Map0862 (a gene uniquely present in M. avium subspecies paratuberculosis) and Map3786 encoded protein solicited the strongest antibody response in infected sheep. The protein encoded by Map2116c showed the weakest antibody response among the animals tested. Although none of the recombinant proteins detected all 11 infected sheep singly, antibodies to Map0862 were detected in 9 of 11 (81%) infected sheep. Furthermore, ovine responses to these selected antigens were assessed temporally over the course of 1 year during which we found a spiking effect rather than an incremental increase of antibody reactivity. This study evaluated multiple M. avium subsp. paratuberculosis recombinant proteins in an ELISA-based format for sheep.
Insights
Researchers evaluated recombinant proteins from Mycobacterium avium subsp. paratuberculosis for diagnosing ovine paratuberculosis using ELISA. Map0862 showed the strongest antibody response in infected sheep, detecting 81% of cases.
Area of Science:
- Veterinary Immunology
- Bacteriology
- Diagnostic Assays
Background:
- Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), affects livestock globally.
- Previous studies identified immunogenic recombinant proteins of MAP in cattle.
- Effective serodiagnostic tools for ovine paratuberculosis are crucial for disease management.
Purpose of the Study:
- To evaluate multiple MAP recombinant proteins for serodiagnosis of ovine paratuberculosis using ELISA.
- To identify specific MAP antigens that elicit strong antibody responses in infected sheep.
- To assess the temporal antibody reactivity to these antigens over one year.
Main Methods:
- An enzyme-linked immunosorbent assay (ELISA) was developed and employed.
- Eighteen purified recombinant proteins of MAP were tested against serum samples from infected and healthy sheep.
- Antibody responses were measured, and temporal reactivity was assessed over 12 months.
Main Results:
- Map0862 and Map3786 encoded proteins elicited the strongest antibody responses in infected sheep.
- Antibodies to Map0862 were detected in 81% (9 of 11) of infected sheep.
- Antibody reactivity showed a spiking pattern over the year rather than a consistent increase.
Conclusions:
- Specific MAP recombinant proteins, particularly Map0862, show promise as diagnostic antigens for ovine paratuberculosis.
- Further refinement of ELISA using these antigens could improve serodiagnosis accuracy.
- Understanding temporal antibody dynamics is important for interpreting diagnostic results.
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