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马尔尼菲青霉蛋白质抗原的蛋白质免疫印迹分析。

Western immunoblot analysis of protein antigens of Penicillium marneffei.

作者信息

Vanittanakom N, Mekaprateep M, Sittisombut N, Supparatpinyo K, Kanjanasthiti P, Nelson K E, Sirisanthana T

机构信息

Department of Microbiology, Faculty of Medicine, Chiang Mai University, Thailand.

出版信息

J Med Vet Mycol. 1997 Mar-Apr;35(2):123-31. doi: 10.1080/02681219780001011.

Abstract

Protein antigens of Penicillium marneffei prepared during the yeast and mould phases of in vitro growth were analyzed by gel electrophoresis and immunoblot assay. More than 20 yeast phase proteins were detected by Coomassie staining; among these, at least 10 reacted with IgG in the pooled sera of 28 AIDS patients with penicilliosis. Four immunogenic proteins of 200, 88, 54 and 50 kDa were produced in large quantity during the deceleration and early stationary phases of growth. When these proteins were reacted with individual sera derived from 33 AIDS patients with penicilliosis, reactivities to the 200, 88, 54 and 50 kDa protein were detected in 72.7, 93.9, 60.6 and 57.6%, respectively. The bands of 88, 54 and 50 kDa gave strong reactions with about a half of serum samples. In one serum derived from an AIDS patient, reactivities to the 54 and 50 kDa proteins could be strongly detected two months before the definite diagnosis by fungal culture. Protein components from the mould form were of lower yield and gave weaker signal in immunoblot analysis. These results indicate that at least two yeast-phase immunoreactive proteins (54 and 50 kDa) are relatively specific to the P. marneffei infection, thereby suggesting its potential for clinical application to the diagnosis of this emerging disease.

摘要

对在体外生长的酵母和霉菌阶段制备的马尔尼菲青霉菌蛋白质抗原进行了凝胶电泳和免疫印迹分析。考马斯亮蓝染色检测到20多种酵母阶段蛋白;其中,至少10种与28例马尔尼菲青霉病艾滋病患者的混合血清中的IgG发生反应。在生长的减速期和早期稳定期大量产生了4种200、88、54和50 kDa的免疫原性蛋白。当这些蛋白与33例马尔尼菲青霉病艾滋病患者的个体血清反应时,对200、88、54和50 kDa蛋白的反应性分别在72.7%、93.9%、60.6%和57.6%中检测到。88、54和50 kDa的条带与大约一半的血清样本发生强烈反应。在一名艾滋病患者的一份血清中,在通过真菌培养明确诊断前两个月就能强烈检测到对54和50 kDa蛋白的反应性。霉菌形式的蛋白质成分产量较低,在免疫印迹分析中信号较弱。这些结果表明,至少两种酵母阶段免疫反应性蛋白(54和50 kDa)对马尔尼菲青霉感染具有相对特异性,从而提示其在临床应用于诊断这种新出现疾病的潜力。

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