Gan Z, Marquardt R R, Abramson D, Clear R M
Department of Animal Science, University of Manitoba, Winnipeg, Canada.
Int J Food Microbiol. 1997 Sep 16;38(2-3):191-200. doi: 10.1016/s0168-1605(97)00108-6.
Immunoassays for the detection of three Fusarium species have been developed. Chickens were immunized with the soluble antigens from within the mycelia (mycelia-soluble antigens) and the exoantigens of Fusarium sporotrichioides, Fusarium poae and Fusarium graminearum and the antigens were characterized using enzyme-linked immunosorbent assays, immunoblotting and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The cross-reactivities of antisera with eleven species of Fusarium, eleven species of fungi from other genera and the buffer-extracts of grain were determined. Antisera to the exoantigens and mycelia-soluble antigens did not cross-react with buffer-extracts of grain. The antisera against the mycelia-soluble antigens tended to cross-react with the antigens from other genera whereas the antisera against the exoantigens were genus specific. The antiserum raised against exoantigens from F. poae was species specific. The molecular weights of the immunodominant antigens from the Fusaria were above 28 kilodaltons. Antigens from Fusaria showed specific bands in SDS-PAGE gel that can be used to produce more specific antibodies. The results suggest that exoantigen immunoassay can be developed to identify Fusarium genus, whereas mycelia-soluble antigens are not suitable for immunoidentification among genus.
已开发出用于检测三种镰刀菌属物种的免疫测定法。用来自拟分枝孢镰刀菌、梨孢镰刀菌和禾谷镰刀菌的菌丝体内部可溶性抗原(菌丝体可溶性抗原)和外抗原对鸡进行免疫,并使用酶联免疫吸附测定、免疫印迹和十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)对这些抗原进行表征。测定了抗血清与11种镰刀菌属物种、11种其他属真菌以及谷物缓冲液提取物的交叉反应性。外抗原和菌丝体可溶性抗原的抗血清与谷物缓冲液提取物无交叉反应。针对菌丝体可溶性抗原的抗血清倾向于与来自其他属的抗原发生交叉反应,而针对外抗原的抗血清具有属特异性。针对梨孢镰刀菌外抗原产生的抗血清具有种特异性。镰刀菌属免疫显性抗原的分子量高于28千道尔顿。镰刀菌属的抗原在SDS-PAGE凝胶中显示出特定条带,可用于产生更特异性的抗体。结果表明,可以开发外抗原免疫测定法来鉴定镰刀菌属,而菌丝体可溶性抗原不适合用于属间的免疫鉴定。