Valentin-Weigand P, Murray C, Moriarty K M
Department of Veterinary Pathology and Public Health, Massey University, Palmerston North, New Zealand.
FEMS Microbiol Lett. 1991 Mar 1;62(2-3):145-8. doi: 10.1016/0378-1097(91)90148-4.
Antibody reactivities in sera from Mycobacterium paratuberculosis (M. ptb) infected and vaccinated sheep were analyzed by enzyme-linked immunosorbent assay (ELISA) and Western (immuno)blotting using a sonicate antigen from M. ptb. Both methods allowed good differentiation between infected/vaccinated animals and noninfected controls. Removal of nonspecific crossreactive antibodies by absorption with a M. phlei sonicate antigen coupled to Sepharose reduced ELISA reactivities of positive sera by 50% and those of noninfected serum by 85%. Immunoblotting analysis revealed that reduction by M. phlei absorption was due to lower reactivities of M. ptb antigens in the range of 30 to 45 kDa. However, one protein with a molecular mass of approx. 27 kDa seemed to be specific for M. ptb since it reacted similarly with nonabsorbed and absorbed serum but not with antibodies which were eluted from M. phlei-Sepharose after absorption. Our findings indicate that M. ptb and M. phlei share a number of common antigens of potential pathogenic importance and that only a smaller part of proteins (i.e. the 27 kDa protein) might be specific for M. ptb.
利用副结核分枝杆菌（M. ptb）的超声破碎抗原，通过酶联免疫吸附测定（ELISA）和蛋白质免疫印迹法分析了感染和接种过疫苗的绵羊血清中的抗体反应性。两种方法都能很好地区分感染/接种动物和未感染对照。用偶联到琼脂糖凝胶上的草分枝杆菌超声破碎抗原进行吸收，去除非特异性交叉反应抗体后，阳性血清的ELISA反应性降低了50%，未感染血清的反应性降低了85%。免疫印迹分析表明，草分枝杆菌吸收导致的反应性降低是由于M. ptb抗原在30至45 kDa范围内的反应性较低。然而，一种分子量约为27 kDa的蛋白质似乎对M. ptb具有特异性，因为它与未吸收和吸收后的血清反应相似，但与吸收后从草分枝杆菌-琼脂糖凝胶上洗脱的抗体不反应。我们的研究结果表明，M. ptb和草分枝杆菌共享一些具有潜在致病重要性的共同抗原，并且只有一小部分蛋白质（即27 kDa蛋白质）可能对M. ptb具有特异性。
