Inzana T J, Todd J, Koch C, Nicolet J
Department of Pathobiology, Virginia Polytechnic Institute and State University, Blacksburg 24061.
Vet Microbiol. 1992 Jun 15;31(4):351-62. doi: 10.1016/0378-1135(92)90127-f.
The cross-reactivity of the purified polysaccharides of Actinobacillus pleuropneumoniae serotypes 1 and 9 were examined using a variety of highly sensitive assays, such as radioimmunoassay, latex agglutination, enzyme-linked immunosorbent assay (ELISA), and immunoblotting. In addition, conventional immunodiffusion was included for comparison. Latex agglutination, utilizing affinity-purified IgG to capsule, was also used to serotype whole cells. Agglutination or precipitation tests (radioimmunoassay, latex agglutination, and immunodiffusion) indicated no cross-reactivity between the capsules of serotypes 1 and 9, and no cross-reactivity between whole cells by latex agglutination. Assays that required binding of the capsule to a solid support (ELISA and immunoblotting) did demonstrate cross-reactions between serotypes 1 and 9 capsules, although reactions with the heterologous serotype were weaker than with the homologous serotype. The cross-reactivity could not be attributed solely to nonspecific factors because similar cross-reactivity did not occur with serotype 5 or 7 capsules by any assay. Reactivity of antisera with homologous or heterologous capsule was reduced, but not completely eliminated, by adsorption with washed, live bacteria of the heterologous serotype. Thus, the assay, as well as the antigen or specificity of the antibody reagent used, may influence the results of A. pleuropneumoniae serotyping or serological tests.
利用多种高灵敏度检测方法,如放射免疫测定、乳胶凝集试验、酶联免疫吸附测定(ELISA)和免疫印迹法,检测了胸膜肺炎放线杆菌1型和9型纯化多糖的交叉反应性。此外,还纳入了传统免疫扩散法进行比较。利用亲和纯化的抗荚膜IgG进行的乳胶凝集试验也用于对全细胞进行血清分型。凝集或沉淀试验(放射免疫测定、乳胶凝集试验和免疫扩散)表明,1型和9型荚膜之间无交叉反应,乳胶凝集试验显示全细胞之间也无交叉反应。需要将荚膜与固相支持物结合的检测方法(ELISA和免疫印迹法)确实显示1型和9型荚膜之间存在交叉反应,尽管与异源血清型的反应比与同源血清型的反应弱。这种交叉反应不能完全归因于非特异性因素,因为任何检测方法对5型或7型荚膜都未出现类似的交叉反应。用异源血清型的洗涤活细菌吸附后,抗血清与同源或异源荚膜的反应性降低,但未完全消除。因此,检测方法以及所用抗体试剂的抗原或特异性可能会影响胸膜肺炎放线杆菌血清分型或血清学检测的结果。