Souza Giliane S, Rodrigues Ana Bárbara F, Gioffré Andrea, Romano Maria I, Carvalho Eulógio C Q, Ventura Thatiana L B, Lasunskaia Elena B
Laboratory of Biology of Recognition, Universidade Estadual do Norte Fluminense, Campos, 28013-602 Rio de Janeiro, Brazil.
Vet Immunol Immunopathol. 2011 Sep 15;143(1-2):75-82. doi: 10.1016/j.vetimm.2011.06.026. Epub 2011 Jun 23.
Comparative genomics of Mycobacterium spp. have revealed conservative genes and respective proteins differently expressed in mycobacteria that could be used as targets for the species-specific immunodiagnostics. The alanine and proline-rich antigen Apa is a mycobacterial protein that present significant variability in primary sequence length and composition between members of M. avium and M. tuberculosis complexes. In this study, the recombinant Apa protein encoded by the MAP1569/ModD gene of M. avium subsp. paratuberculosis (Map) was used to generate a panel of monoclonal antibodies which were shown to recognize the most important veterinary pathogens of the M. avium complex, specifically Map and M. avium subsp. hominissuis, and which did not cross-react with M. bovis or M. tuberculosis. The produced antibodies were demonstrated to be a useful tool for the species-specific immunofluorescence or immunohistochemical detection of Map in experimentally infected cell cultures or intestinal tissues from cattle with bovine paratuberculosis and, additionally, they may be employed for the discrimination of pathogenic M. avium subspecies via Western blotting.
分枝杆菌属的比较基因组学研究揭示了保守基因以及分枝杆菌中差异表达的相应蛋白质，这些可作为种特异性免疫诊断的靶点。富含丙氨酸和脯氨酸的抗原Apa是一种分枝杆菌蛋白，在鸟分枝杆菌和结核分枝杆菌复合体成员之间，其一级序列长度和组成存在显著差异。在本研究中，鸟分枝杆菌副结核亚种（Map）的MAP1569/ModD基因编码的重组Apa蛋白被用于制备一组单克隆抗体，这些抗体被证明能识别鸟分枝杆菌复合体中最重要的兽医病原体，特别是Map和鸟分枝杆菌人亚种，且不与牛分枝杆菌或结核分枝杆菌发生交叉反应。所产生的抗体被证明是一种有用的工具，可用于在实验感染的细胞培养物或患有牛副结核病的牛的肠道组织中对Map进行种特异性免疫荧光或免疫组织化学检测，此外，它们还可用于通过蛋白质印迹法区分致病性鸟分枝杆菌亚种。
