Wiker H G, Harboe M, Nagai S
Institute of Immunology and Rheumatology, University of Oslo, Norway.
J Gen Microbiol. 1991 Apr;137(4):875-84. doi: 10.1099/00221287-137-4-875.
A series of mycobacterial antigens were quantified by immunoelectrophoresis, enzyme-linked immunosorbent assays, or SDS-PAGE with immunoblotting using antisera against purified mycobacterial antigens. The antigens showed a characteristic distribution profile. Some had a marked quantitative dominance in the culture fluid while others had a marked dominance in sonicates of whole washed bacilli. The majority of the antigens tested could thus be located and grouped as either secreted or cytoplasmic in terms of a localization index (LI) which is described. A 5-week-old Mycobacterium tuberculosis culture fluid preparation with a low degree of lysis was valuable in the delineation of localization indexes. The various secreted antigens showed a great span in LI values, from 5 to 1000. This variation may express different degrees of secretion efficiency or differences in tendency to adhere to the bacterial surface. The identification of proteins as extracellular or cytosolic according to their LI values was in agreement for cultures of M. tuberculosis with a low degree of lysis and cultures of M. bovis BCG and M. bovis AN-5 with significant lysis of the bacterial cells.
通过免疫电泳、酶联免疫吸附测定或十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)结合使用针对纯化分枝杆菌抗原的抗血清进行免疫印迹,对一系列分枝杆菌抗原进行了定量分析。这些抗原呈现出特征性的分布图谱。有些抗原在培养液中具有明显的数量优势,而另一些则在整个洗涤后的杆菌超声裂解物中具有明显优势。因此,根据所描述的定位指数(LI),所测试的大多数抗原可定位并归类为分泌型或胞质型。一种5周龄、裂解程度低的结核分枝杆菌培养液制剂在确定定位指数方面很有价值。各种分泌型抗原的LI值跨度很大,从5到1000。这种变化可能表示不同程度的分泌效率或附着于细菌表面倾向的差异。根据LI值将蛋白质鉴定为细胞外或胞质内,对于裂解程度低的结核分枝杆菌培养物以及细菌细胞有明显裂解的牛分枝杆菌卡介苗和牛分枝杆菌AN-5培养物来说是一致的。