Gandra R F, Melo T A, Matsumoto F E, Pires M F C, Croce J, Gambale W, Paula C R
Departamento de Microbiologia, Universidade de São Paulo, Brasil.
Mycopathologia. 2002;155(4):183-9. doi: 10.1023/a:1021181711225.
Crude extracts of the lipophilic yeast Malassezia furfur were obtained from 2, 6, 10 and 28 day old cultures. The in vitro cultivation periods corresponded, respectively, to the lag phase, middle of the log phase, end of log phase and the decline phase of the growth curve, which was based on viable cell counts obtained with a fluorescent viability test. Biochemical analyses showed that the protein and carbohydrate contents were greater in day 10 extracts. Seventy patients with different allergic manifestations and 30 healthy volunteers were skin prick tested using the extracts. Of these, thirteen (18.57%) patients gave positive responses. SDS PAGE gradient electrophoretic profiles of the preparations indicated that the 28 day extracts contained the greatest number of protein bands with molecular weights ranging mostly between 30 and 94 kDa. Immunoblots incubated with individual patient sera showed that four IgE binding M. furfur allergens of approximately 88, 61, 52 and 39 kDa were present in the 28 day extracts. The components identified could be used for detecting IgE mediated responses to M. furfur among individuals affected with different allergic conditions.
亲脂性酵母糠秕马拉色菌的粗提物取自培养2天、6天、10天和28天的培养物。体外培养期分别对应生长曲线的延迟期、对数期中期、对数期末期和衰退期，该生长曲线基于通过荧光活力测试获得的活细胞计数。生化分析表明，第10天提取物中的蛋白质和碳水化合物含量更高。使用这些提取物对70名有不同过敏表现的患者和30名健康志愿者进行了皮肤点刺试验。其中，13名（18.57%）患者呈阳性反应。制剂的SDS-PAGE梯度电泳图谱表明，28天提取物中蛋白质条带数量最多，分子量大多在30至94 kDa之间。与个体患者血清孵育的免疫印迹显示，28天提取物中存在四种约88、61、52和39 kDa的IgE结合糠秕马拉色菌过敏原。所鉴定的成分可用于检测不同过敏状况个体中对糠秕马拉色菌的IgE介导反应。
