Bannantine John P, Rosu Valentina, Zanetti Stefania, Rocca Stefano, Ahmed Niyaz, Sechi Leonardo A
National Animal Disease Center, USDA-Agricultural Research Service, Ames, IA 50010, USA.
Vet Immunol Immunopathol. 2008 Mar 15;122(1-2):116-25. doi: 10.1016/j.vetimm.2007.10.020. Epub 2007 Nov 9.
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.
数年来一直在探索改进酶联免疫吸附测定（ELISA）试验以检测副结核分枝杆菌的方法。此前，已发现副结核分枝杆菌的某些重组蛋白对患有副结核病的牛具有免疫原性。在本研究中，使用18种纯化的重组蛋白以ELISA形式评估感染和健康绵羊的抗体反应，用于绵羊副结核病的血清学诊断。这些选定的重组蛋白代表副结核分枝杆菌的热休克蛋白、假定蛋白和细胞表面蛋白。然而，Map0862（副结核分枝杆菌中独特存在的一个基因）和Map3786编码的蛋白在感染绵羊中引发了最强的抗体反应。Map2116c编码的蛋白在受试动物中显示出最弱的抗体反应。尽管没有一种重组蛋白能单独检测出所有11只感染绵羊，但在11只感染绵羊中有9只（81%）检测到了针对Map0862的抗体。此外，在1年的时间里对绵羊对这些选定抗原的反应进行了时间上的评估，在此期间我们发现了一种峰值效应，而不是抗体反应性的逐渐增加。本研究以ELISA形式评估了多种副结核分枝杆菌重组蛋白对绵羊的作用。
