马尼菲青霉蛋白抗原的免疫印迹分析。

N Vanittanakom, M Mekaprateep, N Sittisombut, K Supparatpinyo, P Kanjanasthiti, K E Nelson, T Sirisanthana
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引用次数: 29

摘要

采用凝胶电泳和免疫印迹法对体外生长酵母期和霉菌期制备的马尼菲青霉蛋白抗原进行了分析。考马斯氏染色法检测到20多个酵母相蛋白;其中,28例艾滋病青霉菌病患者血清中至少有10例与IgG反应。在生长减速和早期稳定期大量产生了200、88、54和50 kDa的4种免疫原性蛋白。当这些蛋白与33例艾滋病青霉菌病患者的血清反应时,检测到200、88、54和50 kDa蛋白的反应率分别为72.7%、93.9%、60.6%和57.6%。88、54和50 kDa的条带对半数左右的血清样品有强烈反应。在一名艾滋病患者的血清中,通过真菌培养,可以在确诊前两个月强烈检测到54和50 kDa蛋白的反应。从霉菌中提取的蛋白质成分产量较低,免疫印迹分析信号较弱。这些结果表明,至少有两种酵母期免疫反应蛋白(54和50 kDa)对P. marneffi感染具有相对特异性,从而表明其在诊断这种新发疾病方面具有临床应用潜力。
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Western immunoblot analysis of protein antigens of Penicillium marneffei.

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.

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