Kurup V P, Greenberger P A, Fink J N
Research Service, Veterans Administration Medical Center, Milwaukee, Wisconsin.
J Clin Microbiol. 1989 Jun;27(6):1312-6. doi: 10.1128/jcm.27.6.1312-1316.1989.
Sera from patients with allergic bronchopulmonary aspergillosis (ABPA) or aspergilloma and normal sera were analyzed for specific antibodies by Western (immuno-) blotting with Aspergillus fumigatus antigens transferred electrophoretically onto polyvinylidene difluoride membranes. Western blot analysis demonstrated consistent reactivity of low-molecular-weight A. fumigatus antigens against ABPA sera but not against uncomplicated aspergilloma or normal sera. None of these low-molecular-weight components had any lectin-binding activity. Sera from patients with aspergilloma, however, frequently reacted with high-molecular-weight components of A. fumigatus. The majority of these high-molecular-weight antigenic components demonstrated concanavalin A-binding activity. The low-molecular-weight bands were discernible in Western blots with sera from all ABPA patients irrespective of disease activities, such as relapse, flare, or treatment. Antibodies detected by methods such as immunodiffusion or enzyme-linked immunosorbent assays demonstrated total antibody responses to most or all antigenic components, while Western blots demonstrated the reactivities of the individual components with the specific antibodies. Western blot analysis thus provided more information for immunodiagnosis of ABPA than other methods, especially when only crude antigens were available.
采用电泳法将烟曲霉抗原转移至聚偏二氟乙烯膜上,通过蛋白质免疫印迹法对过敏性支气管肺曲霉病(ABPA)患者、曲霉肿患者的血清以及正常血清进行特异性抗体分析。蛋白质免疫印迹分析表明,低分子量烟曲霉抗原与ABPA患者血清有持续反应,但与单纯曲霉肿患者血清或正常血清无反应。这些低分子量成分均无凝集素结合活性。然而,曲霉肿患者的血清经常与烟曲霉的高分子量成分发生反应。这些高分子量抗原成分大多具有伴刀豆球蛋白A结合活性。无论疾病处于复发、发作或治疗等何种活动状态,所有ABPA患者血清的蛋白质免疫印迹中均可辨别出低分子量条带。免疫扩散或酶联免疫吸附测定等方法检测到的抗体显示了对大多数或所有抗原成分的总抗体反应,而蛋白质免疫印迹则显示了各个成分与特异性抗体的反应性。因此,与其他方法相比,蛋白质免疫印迹分析为ABPA的免疫诊断提供了更多信息,尤其是在只有粗抗原可用时。